Compositions and methods relating to lung specific genes

ABSTRACT

The invention relates to LSG polypeptides, polynucleotides encoding the polypeptides, methods for producing the polypeptides, in particular by expressing the polynucleotides, and agonists and antagonists of the polypeptides. The invention further relates to methods for utilizing such polynucleotides, polypeptides, agonists and antagonists for applications, which relate, in part, to research, diagnostic and clinical arts.

INTRODUCTION

[0001] This application claims the benefit of priority from U.S.Provisional Application Serial No. 60/219,834, filed Jul. 21, 2000,which is herein incorporated in its entirety.

FIELD OF THE INVENTION

[0002] The present invention relates to newly identified nucleic acidsand polypeptides present in normal and neoplastic lung cells, includingfragments, variants and derivatives of the nucleic acids andpolypeptides. The present invention also relates to antibodies to thepolypeptides of the invention, as well as agonists and antagonists ofthe polypeptides of the invention. The invention also relates tocompositions comprising the nucleic acids, polypeptides, antibodies,variants, derivatives, agonists and antagonists of the invention andmethods for the use of these compositions. These uses includeidentifying, diagnosing, monitoring, staging, imaging and treating lungcancer and non-cancerous disease states in lung, identifying lungtissue, monitoring and modifying lung embryonic development anddifferentiation, and identifying and/or designing agonists andantagonists of polypeptides of the invention. The uses also include genetherapy, production of transgenic animals and cells, and production ofengineered lung tissue for treatment and research.

BACKGROUND OF THE INVENTION

[0003] Throughout the last hundred years, the incidence of lung cancerhas steadily increased, so much so that now in many countries, it is themost common cancers. In fact, lung cancer is the second most prevalenttype of cancer for both men and women in the United States and is themost common cause of cancer death in both sexes. Lung cancer deaths haveincreased ten-fold in both men and women since 1930, primarily due to anincrease in cigarette smoking, but also due to an increased exposure toarsenic, asbestos, chromates, chloromethyl ethers, nickel, polycyclicaromatic hydrocarbons and other agents. See Scott, Lung Cancer: A Guideto Diagnosis and Treatment, Addicus Books (2000) and Alberg et al., inKane et al. (eds.) Biology of Lung Cancer, pp. 11-52, Marcel Dekker,Inc. (1998). Lung cancer may result from a primary tumor originating inthe lung or a secondary tumor which has spread from another organ suchas the bowel or breast. Although there are over a dozen types of lungcancer, over 90% fall into two categories: small cell lung cancer (SCLC)and non-small cell lung cancer (NSCLC). See Scott, supra. About 20-25%of all lung cancers are characterized as SCLC, while 70-80% arediagnosed as NSCLC. Id. A rare type of lung cancer is mesothelioma,which is generally caused by exposure to asbestos, and which affects thepleura of the lung. Lung cancer is usually diagnosed or screened for bychest x-ray, CAT scans, PET scans, or by sputum cytology. A diagnosis oflung cancer is usually confirmed by biopsy of the tissue. Id.

[0004] SCLC tumors are highly metastatic and grow quickly. By the time apatient has been diagnosed with SCLC, the cancer has usually alreadyspread to other parts of the body, including lymph nodes, adrenals,liver, bone, brain and bone marrow. See Scott, supra; Van Houtte et al.(eds.), Progress and Perspective in the Treatment of Lung Cancer,Springer-Verlag (1999). Because the disease has usually spread to suchan extent that surgery is not an option, the current treatment of choiceis chemotherapy plus chest irradiation. See Van Houtte, supra. The stageof disease is a principal predictor of long-term survival.

[0005] Less than 5% of patients with extensive disease that has spreadbeyond one lung and surrounding lymph nodes, live longer than two years.Id. However, the probability of five-year survival is three to fourtimes higher if the disease is diagnosed and treated when it is still ina limited stage, i.e., not having spread beyond one lung.

[0006] NSCLC is generally divided into three types: squamous cellcarcinoma, adenocarcinoma and large cell carcinoma. Both squamous cellcancer and adenocarcinoma develop from the cells that line the airways;however, adenocarcinoma develops from the goblet cells that producemucus. Large cell lung cancer has been thus named because the cells looklarge and rounded when viewed microscopically, and generally areconsidered relatively undifferentiated. See Yesner, Atlas of LungCancer, Lippincott-Raven (1998).

[0007] Secondary lung cancer is a cancer initiated elsewhere in the bodythat has spread to the lungs. Cancers that metastasize to the lunginclude, but are not limited to, breast cancer, melanoma, colon cancerand Hodgkin's lymphoma. Treatment for secondary lung cancer may dependupon the source of the original cancer. In other words, a lung cancerthat originated from breast cancer may be more responsive to breastcancer treatments and a lung cancer that originated from the coloncancer may be more responsive to colon cancer treatments.

[0008] The stage of a cancer indicates how far it has spread and is animportant indicator of the prognosis. In addition, staging is importantbecause treatment is often decided according to the stage of a cancer.SCLC is divided into two stages: limited disease, i.e., cancer that canonly be seen in one lung and in nearby lymph nodes; and extensivedisease, i.e., cancer that has spread outside the lung to the chest orto other parts of the body. For most patients with SCLC, the disease hasalready progressed to lymph nodes or elsewhere in the body at the timeof diagnosis. See Scott, supra. Even if spreading is not apparent on thescans, it is likely that some cancer cells may have spread away andtraveled through the bloodstream or lymph system. In general,chemotherapy with or without radiotherapy is often the preferredtreatment. The initial scans and tests done at first will be used laterto see how well a patient is responding to treatment.

[0009] In contrast, non-small cell cancer may be divided into fourstages. Stage I is highly localized cancer with no cancer in the lymphnodes. Stage II cancer has spread to the lymph nodes at the top of theaffected lung. Stage III cancer has spread near to where the cancerstarted. This can be to the chest wall, the covering of the lung(pleura), the middle of the chest (mediastinum) or other lymph nodes.Stage IV cancer has spread to another part of the body. Stage I-IIIcancer is usually treated with surgery, with or without chemotherapy.Stage IV cancer is usually treated with chemotherapy and/or palliativecare.

[0010] A number of chromosomal and genetic abnormalities have beenobserved in lung cancer. In NSCLC, chromosomal aberrations have beendescribed on 3p, 9p, 11p, 15p and 17p, and chromosomal deletions havebeen seen on chromosomes 7, 11, 13 and 19. See Skarin (ed.),Multimodality Treatment of Lung Cancer, Marcel Dekker, Inc. (2000);Gemmill et al., pp. 465-502, in Kane, supra; Bailey-Wilson et al., pp.53-98, in Kane, supra. Chromosomal abnormalities have been described on1p, 3p, 5q, 6q, 8q, 13q and 17p in SCLC. Id. In addition, the loss ofthe short arm of chromosome 3p has also been seen in greater than 90% ofSCLC tumors and approximately 50% of NSCLC tumors. Id.

[0011] A number of oncogenes and tumor suppressor genes have beenimplicated in lung cancer. See Mabry, pp. 391-412, in Kane, supra andSclafani et al., pp. 295-316, in Kane, supra. In both SCLC and NSCLC,the p53 tumor suppressor gene is mutated in over 50% of lung cancers.See Yesner, supra. Another tumor suppressor gene, FHIT, which is foundon chromosome 3p, is mutated by tobacco smoke. Id.;

[0012] Skarin, supra. In addition, more than 95% of SCLCs andapproximately 20-60% of NSCLCs have an absent or abnormal retinoblastoma(Rb) protein, another tumor suppressor gene. The ras oncogene(particularly K-ras) is mutated in 20-30% of NSCLC specimens and thec-erbB2 oncogene is expressed in 18% of stage 2 NSCLC and 60% of stage 4NSCLC specimens. See Van Houtte, supra. Other tumor suppressor genesthat are found in a region of chromosome 9, specifically in the regionof 9p21, are deleted in many cancer cells, including p16^(INK4A) andp15^(INK4B). See Bailey-Wilson, supra; Sclafani et al., supra. Thesetumor suppressor genes may also be implicated in lung cancerpathogenesis.

[0013] In addition, many lung cancer cells produce growth factors thatmay act in an autocrine fashion on lung cancer cells. See Siegfried etal., pp. 317-336, in Kane, supra; Moody, pp. 337-370, in Kane, supra andHeasley et al., 371-390, in Kane, supra. In SCLC, many tumor cellsproduce gastrin-releasing peptide (GRP), which is a proliferative growthfactor for these cells. See Skarin, supra. Many NSCLC tumors expressepidermal growth factor (EGF) receptors, allowing NSCLC cells toproliferate in response to EGF. Insulin-like growth factor (IGF-I) iselevated in greater than 95% of SCLC and greater than 80% of NSCLCtumors; it is thought to function as an autocrine growth factor. Id.Finally, stem cell factor (SCF, also known as steel factor or kitligand) and c-Kit (a proto-oncoprotein tyrosine kinase receptor for SCF)are both expressed at high levels in SCLC, and thus may form anautocrine loop that increases proliferation. Id.

[0014] Although the majority of lung cancer cases are attributable tocigarette smoking, most smokers do not develop lung cancer.Epidemiological evidence has suggested that susceptibility to lungcancer may be inherited in a Mendelian fashion, and thus have aninherited genetic component. Bailey-Wilson, supra. Thus, it is thoughtthat certain allelic variants at some genetic loci may affectsusceptibility to lung cancer. Id. One way to identify which allelicvariants are likely to be involved in lung cancer susceptibility, aswell as susceptibility to other diseases, is to look at allelic variantsof genes that are highly expressed in lung.

[0015] The lung is also susceptible to a number of other debilitatingdiseases, including, without limitation, emphysema, pneumonia, cysticfibrosis and asthma. See Stockley (ed.), Molecular Biology of the Lung,Volume I: Emphysema and Infection, Birkhauser Verlag (1999), hereafterStockley I, and Stockley (ed.), Molecular Biology of the Lung, VolumeII: Asthma and Cancer, Birkhauser Verlag (1999), hereafter Stockley II.The cause of many these disorders is still not well understood and thereare few, if any, good treatment options for many of these noncancerouslung disorders. Thus, there remains a need to understand variousnoncancerous lung disorders and to identify treatments for thesediseases.

[0016] In yet another aspect, the development and differentiation of thelung tissue is important during embryonic development. All of theepithelial cells of the respiratory tract, including those of the lungand bronchi, are derived from the primitive endodermal cells that linethe embryonic outpouching. See Yesner, supra. During embryonicdevelopment, multipotent endodermal stem cells differentiate into manydifferent types of specialized cells, which include ciliated cells formoving inhaled particles, goblet cells for producing mucus, Kulchitsky'scells for endocrine function, and Clara cells and type II pneumocytesfor secreting surfactant protein. Id. Improper development anddifferentiation may cause respiratory disorders and distress in infants,particularly in premature infants, whose lungs cannot produce sufficientsurfactant when they are born. Further, some lung cancer cells,particularly small cell carcinomas, appear multipotent, and canspontaneously differentiate into a number of cell types, including smallcell carcinoma, adenocarcinoma and squamous cell carcinoma. Id. Thus, abetter understanding of lung development and differentiation may helpfacilitate understanding of lung cancer initiation and progression.

[0017] Accordingly, there is a great need for more sensitive andaccurate methods for predicting whether a person is likely to developlung cancer, for diagnosing lung cancer, for monitoring the progressionof the disease, for staging the lung cancer, for determining whether thelung cancer has metastasized and for imaging the lung cancer. There isalso a need for better treatment of lung cancer. Further, there is alsoa great need for diagnosing and treating noncancerous lung disorderssuch as emphysema, pneumonia, lung infection, pulmonary fibrosis, cysticfibrosis and asthma. There is also a need for compositions and methodsof using them that can be used to identify lung tissue for forensicpurposes and for determining whether a particular cell or tissueexhibits lung-specific characteristics.

[0018] In the present invention, methods are provided for detecting,diagnosing, monitoring, staging, prognosticating, imaging and treatinglung cancer via lung specific genes referred to herein as LSGs. Forpurposes of the present invention, LSG refers, among other things, tonative protein expressed by the gene comprising a polynucleotidesequence of SEQ ID NO:1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,16, 17, 18, 19, 20, 21, or 22 or a contig of SEQ ID NO: 19 or 21 asdepicted in SEQ ID NO: 37, or 38, respectively. By “LSG” it is alsomeant herein polynucleotides which, due to degeneracy in genetic coding,comprise variations in nucleotide sequence as compared to SEQ ID NO: 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21,22, 37, 38, 39 or 40 but which still encode the same polypeptide.Exemplary amino acid sequences for LSG polypeptides are set forth in SEQID NO: 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54,55 and 56. In the alternative, what is meant by LSG as used herein,means the native mRNA encoded by the gene comprising the polynucleotidesequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14,15, 16, 17, 18, 19, 20, 21 or 22, levels of the gene comprising thepolynucleotide sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11,12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or 22 or levels of apolynucleotide which is capable of hybridizing under stringentconditions to the antisense sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7,8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 37, or 38.

[0019] Other objects, features, advantages and aspects of the presentinvention will become apparent to those of skill in the art from thefollowing description. It should be understood, however, that thefollowing description and the specific examples, while indicatingpreferred embodiments of the invention are given by way of illustrationonly. Various changes and modifications within the spirit and scope ofthe disclosed invention will become readily apparent to those skilled inthe art from reading the following description and from reading theother parts of the present disclosure.

SUMMARY OF THE INVENTION

[0020] Toward these ends, and others, it is an object of the presentinvention to provide LSGs comprising a polynucleotide of SEQ ID NO: 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21,22, 37 or 38 a protein expressed by a polynucleotide of SEQ ID NO: 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22,37 or 38 or a variant thereof which expresses the protein; or apolynucleotide which is capable of hybridizing under stringentconditions to the antisense sequence of SEQ ID NO:1, 2, 3, 4, 5, 6, 7,8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 37 or 38.Exemplary LSG polypeptides of the present invention are depicted in SEQDI NO: 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54,55 or 56.

[0021] It is another object of the present invention to provide a methodfor diagnosing the presence of lung cancer by analyzing for changes inlevels of LSG in cells, tissues or bodily fluids compared with levels ofLSG in preferably the same cells, tissues, or bodily fluid type of anormal human control, wherein a change in levels of LSG in the patientversus the normal human control is associated with lung cancer.

[0022] Further provided is a method of diagnosing metastatic lung cancerin a patient having lung cancer which is not known to have metastasizedby identifying a human patient suspected of having lung cancer that hasmetastasized; analyzing a sample of cells, tissues, or bodily fluid fromsuch patient for LSG; comparing the LSG levels in such cells, tissues,or bodily fluid with levels of LSG in preferably the same cells,tissues, or bodily fluid type of a normal human control, wherein anincrease in LSG levels in the patient versus the normal human control isassociated with lung cancer which has metastasized.

[0023] Also provided by the invention is a method of staging lung cancerin a human which has such cancer by identifying a human patient havingsuch cancer; analyzing a sample of cells, tissues, or bodily fluid fromsuch patient for LSG; comparing LSG levels in such cells, tissues, orbodily fluid with levels of LSG in preferably the same cells, tissues,or bodily fluid type of a normal human control sample, wherein anincrease in LSG levels in the patient versus the normal human control isassociated with a cancer which is progressing and a decrease in thelevels of LSG is associated with a cancer which is regressing or inremission.

[0024] Further provided is a method of monitoring lung cancer in a humanhaving such cancer for the onset of metastasis. The method comprisesidentifying a human patient having such cancer that is not known to havemetastasized; periodically analyzing a sample of cells, tissues, orbodily fluid from such patient for LSG; comparing the LSG levels in suchcells, tissue, or bodily fluid with levels of LSG in preferably the samecells, tissues, or bodily fluid type of a normal human control sample,wherein an increase in LSG levels in the patient versus the normal humancontrol is associated with a cancer which has metastasized.

[0025] Further provided is a method of monitoring the change in stage oflung cancer in a human having such cancer by looking at levels of LSG ina human having such cancer. The method comprises identifying a humanpatient having such cancer; periodically analyzing a sample of cells,tissues, or bodily fluid from such patient for LSG; comparing the LSGlevels in such cells, tissue, or bodily fluid with levels of LSG inpreferably the same cells, tissues, or bodily fluid type of a normalhuman control sample, wherein an increase in LSG levels in the patientversus the normal human control is associated with a cancer which isprogressing and a decrease in the levels of LSG is associated with acancer which is regressing or in remission.

[0026] Further provided are methods of designing new therapeutic agentstargeted to a LSG for use in imaging and treating lung cancer. Forexample, in one embodiment, therapeutic agents such as antibodiestargeted against LSG or fragments of such antibodies can be used totreat, detect or image localization of LSG in a patient for the purposeof detecting or diagnosing a disease or condition. In this embodiment,an increase in the amount of labeled antibody detected as compared tonormal tissue would be indicative of tumor metastases or growth. Suchantibodies can be polyclonal, monoclonal, or omniclonal or prepared bymolecular biology techniques. The term “antibody”, as used herein andthroughout the instant specification is also meant to include aptamersand single-stranded oligonucleotides such as those derived from an invitro evolution protocol referred to as SELEX and well known to thoseskilled in the art. Antibodies can be labeled with a variety ofdetectable and therapeutic labels including, but not limited to,radioisotopes and paramagnetic metals. Therapeutic agents such as smallmolecules and antibodies which decrease the concentration and/oractivity of LSG can also be used in the treatment of diseasescharacterized by overexpression of LSG. Such agents can be readilyidentified in accordance with teachings herein.

[0027] Other objects, features, advantages and aspects of the presentinvention will become apparent to those of skill in the art from thefollowing description. It should be understood, however, that thefollowing description and the specific examples, while indicatingpreferred embodiments of the invention, are given by way of illustrationonly. Various changes and modifications within the spirit and scope ofthe disclosed invention will become readily apparent to those skilled inthe art from reading the following description and from reading theother parts of the present disclosure.

[0028] Glossary

[0029] The following illustrative explanations are provided tofacilitate understanding of certain terms used frequently herein,particularly in the examples. The explanations are provided as aconvenience and are not limitative of the invention.

[0030] ISOLATED means altered “by the hand of man” from its naturalstate; i.e., that, if it occurs in nature, it has been changed orremoved from its original environment, or both.

[0031] For example, a naturally occurring polynucleotide or apolypeptide naturally present in a living animal in its natural state isnot “isolated,” but the same polynucleotide or polypeptide separatedfrom the coexisting materials of its natural state is “isolated”, as theterm is employed herein. For example, with respect to polynucleotides,the term isolated means that it is separated from the chromosome andcell in which it naturally occurs.

[0032] As part of or following isolation, such polynucleotides can bejoined to other polynucleotides, such as DNAs, for mutagenesis, to formfusion proteins, and for propagation or expression in a host, forinstance. The isolated polynucleotides, alone or joined to otherpolynucleotides such as vectors, can be introduced into host cells, inculture or in whole organisms. When introduced into host cells inculture or in whole organisms, such DNAs still would be isolated, as theterm is used herein, because they would not be in their naturallyoccurring form or environment. Similarly, the polynucleotides andpolypeptides may occur in a composition, such as media formulations,solutions for introduction of polynucleotides or polypeptides, forexample, into cells, compositions or solutions for chemical or enzymaticreactions, for instance, which are not naturally occurring compositions,and, therein remain isolated polynucleotides or polypeptides within themeaning of that term as it is employed herein.

[0033] OLIGONUCLEOTIDE(S) refers to relatively short polynucleotides.Often the term refers to single-stranded deoxyribonucleotides, but itcan refer as well to single-or double-stranded ribonucleotides, RNA:DNAhybrids and double-stranded DNAs, among others. Oligonucleotides, suchas single-stranded DNA probe oligonucleotides, often are synthesized bychemical methods, such as those implemented on automated oligonucleotidesynthesizers. However, oligonucleotides can be made by a variety ofother methods, including in vitro recombinant DNA-mediated techniquesand by expression of DNAs in cells and organisms.

[0034] Initially, chemically synthesized DNAs typically are obtainedwithout a 5′ phosphate. The 5′ ends of such oligonucleotides are notsubstrates for phosphodiester bond formation by ligation reactions thatemploy DNA ligases typically used to form recombinant DNA molecules.Where ligation of such oligonucleotides is desired, a phosphate can beadded by standard techniques, such as those that employ a kinase andATP.

[0035] The 3′ end of a chemically synthesized oligonucleotide generallyhas a free hydroxyl group and, in the presence of a ligase such as T4DNA ligase, readily will form a phosphodiester bond with a 5′ phosphateof another polynucleotide, such as another oligonucleotide. As is wellknown, this reaction can be prevented selectively, where desired, byremoving the 5′ phosphates of the other polynucleotide(s) prior toligation.

[0036] POLYNUCLEOTIDE(S) generally refers to any polyribonucleotide orpolydeoxribonucleotide and is inclusive of unmodified RNA or DNA as wellas modified RNA or DNA. Thus, for instance, polynucleotides as usedherein refers to, among other things, single- and double-stranded DNA,DNA that is a mixture of single- and double-stranded regions, single-and double-stranded RNA, and RNA that is mixture of single- anddouble-stranded regions, hybrid molecules comprising DNA and RNA thatmay be single-stranded or, more typically, double-stranded or a mixtureof single- and double-stranded regions. In addition, polynucleotide, asused herein, refers to triple-stranded regions comprising RNA or DNA orboth RNA and DNA. The strands in such regions may be from the samemolecule or from different molecules. The regions may include all of oneor more of the molecules, but more typically involve only a region ofsome of the molecules. One of the molecules of a triple-helical regionoften is an oligonucleotide.

[0037] As used herein, the term polynucleotide is also inclusive of DNAsor RNAs as described above that contain one or more modified bases.Thus, DNAs or RNAs with backbones modified for stability or for otherreasons are “polynucleotides” as that term is intended herein. Moreover,DNAs or RNAs comprising unusual bases, such as inosine, or modifiedbases, such as tritylated bases, to name just two examples, arepolynucleotides as the term is used herein.

[0038] It will be appreciated that a great variety of modifications havebeen made to DNA and RNA that serve many useful purposes known to thoseof skill in the art. The term polynucleotide as it is employed hereinembraces such chemically, enzymatically or metabolically modified formsof polynucleotides, as well as chemical forms of DNA and RNAcharacteristic of viruses and cells, including simple and complex cells,inter aliai.

[0039] POLYPEPTIDES, as used herein, includes all polypeptides asdescribed below. The basic structure of polypeptides is well known andhas been described in innumerable textbooks and other publications inthe art.

[0040] In this context, the term is used herein to refer to any peptideor protein comprising two or more amino acids joined to each other in alinear chain by peptide bonds. As used herein, the term refers to bothshort chains, which also commonly are referred to in the art aspeptides, oligopeptides and oligomers, for example, and to longerchains, which generally are referred to in the art as proteins, of whichthere are many types. It will be appreciated that polypeptides oftencontain amino acids other than the 20 amino acids commonly referred toas the naturally occurring amino acids, and that many amino acids,including the terminal amino acids, may be modified in a givenpolypeptide, either by natural processes such as processing and otherpost-translational modifications, or by chemical modification techniqueswhich are well known to the art. Even the common modifications thatoccur naturally in polypeptides are too numerous to list exhaustivelyhere, but they are well described in basic texts and in more detailedmonographs, as well as in a voluminous research literature, and they arewell known to those of skill in the art.

[0041] Modifications which may be present in polypeptides of the presentinvention include, to name an illustrative few, acetylation, acylation,ADP-ribosylation, amidation, covalent attachment of flavin, covalentattachment of a heme moiety, covalent attachment of a nucleotide ornucleotide derivative, covalent attachment of a lipid or lipidderivative, covalent attachment of phosphotidylinositol, cross-linking,cyclization, disulfide bond formation, demethylation, formation ofcovalent cross-links, formation of cystine, formation of pyroglutamate,formylation, gamma-carboxylation, glycosylation, GPI anchor formation,hydroxylation, iodination, methylation, myristoylation, oxidation,proteolytic processing, phosphorylation, prenylation, racemization,selenoylation, sulfation, transfer-RNA mediated addition of amino acidsto proteins such as arginylation, and ubiquitination.

[0042] Such modifications are well known to those of skill and have beendescribed in great detail in the scientific literature. Severalparticularly common modifications including, but not limited to,glycosylation, lipid attachment, sulfation, gamma-carboxylation ofglutamic acid residues, hydroxylation and ADP-ribosylation are describedin most basic texts, such as, for instance PROTEINS STRUCTURE ANDMOLECULAR PROPERTIES, 2nd Ed., T. E. Creighton, W. H. Freeman andCompany, New York (1993). Many detailed reviews are available on thissubject, such as, for example, those provided by Wold, F.,Posttranslational Protein Modifications: Perspectives and Prospects,pgs. 1-12 in POSTTRANSLATIONAL COVALENT MODIFICATION OF PROTEINS, B. C.Johnson, Ed., Academic Press, New York (1983); Seifter et al., Analysisfor protein modifications and nonprotein cofactors, Meth. Enzymol. 182:626-646 (1990) and Rattan et al., Protein Synthesis: PosttranslationalModifications and Aging, Ann. N.Y. Acad. Sci. 663: 48-62 (1992).

[0043] It will be appreciated that the polypeptides of the presentinvention are not always entirely linear. Instead, polypeptides may bebranched as a result of ubiquitination, and they may be circular, withor without branching, generally as a result of posttranslation eventsincluding natural processing event and events brought about by humanmanipulation which do not occur naturally. Circular, branched andbranched circular polypeptides may be synthesized by non-translationnatural processes and by entirely synthetic methods, as well.

[0044] Modifications can occur anywhere in a polypeptide, including thepeptide backbone, the amino acid side-chains and the amino or carboxyltermini. In fact, blockage of the amino and/or carboxyl group in apolypeptide by a covalent modification is common in naturally occurringand synthetic polypeptides and such modifications may be present inpolypeptides of the present invention, as well. For instance, the aminoterminal residue of polypeptides made in E. coli, prior to proteolyticprocessing, almost invariably will be N-formylmethionine.

[0045] The modifications that occur in a polypeptide often will be afunction of how it is made. For polypeptides made by expressing a clonedgene in a host, for instance, the nature and extent of themodifications, in large part, will be determined by the host cellposttranslational modification capacity and the modification signalspresent in the polypeptide amino acid sequence. For instance, as is wellknown, glycosylation often does not occur in bacterial hosts such as E.coli. Accordingly, when glycosylation is desired, a polypeptide can beexpressed in a glycosylating host, generally a eukaryotic cell. Insectcells often carry out the same posttranslational glycosylations asmammalian cells. Thus, insect cell expression systems have beendeveloped to express efficiently mammalian proteins having nativepatterns of glycosylation, inter alia. Similar considerations apply toother modifications.

[0046] It will be appreciated that the same type of modification may bepresent in the same or varying degrees at several sites in a givenpolypeptide. Also, a given polypeptide may contain many types ofmodifications.

[0047] In general, as used herein, the term polypeptide encompasses allsuch modifications, particularly those that are present in polypeptidessynthesized by expressing a polynucleotide in a host cell.

[0048] VARIANT(S) of polynucleotides or polypeptides, as the term isused herein, are polynucleotides or polypeptides that differ from areference polynucleotide or polypeptide, respectively.

[0049] With respect to variant polynucleotides, differences aregenerally limited so that the nucleotide sequences of the reference andthe variant are closely similar overall and, in many regions, identical.Thus, changes in the nucleotide sequence of the variant may be silent.That is, they may not alter the amino acids encoded by thepolynucleotide. Where alterations are limited to silent changes of thistype a variant will encode a polypeptide with the same amino acidsequence as the reference. Alternatively, changes in the nucleotidesequence of the variant may alter the amino acid sequence of apolypeptide encoded by the reference polynucleotide. Such nucleotidechanges may result in amino acid substitutions, additions, deletions,fusions and truncations in the polypeptide encoded by the referencesequence.

[0050] With respect to variant polypeptides, differences are generallylimited so that the sequences of the reference and the variant areclosely similar overall and, in many region, identical. For example, avariant and reference polypeptide may differ in amino acid sequence byone or more substitutions, additions, deletions, fusions andtruncations, which may be present in any combination.

[0051] RECEPTOR MOLECULE, as used herein, refers to molecules which bindor interact specifically with LSG polypeptides of the present inventionand is inclusive not only of classic receptors, which are preferred, butalso other molecules that specifically bind to or interact withpolypeptides of the invention (which also may be referred to as “bindingmolecules” and “interaction molecules,” respectively and as “LSG bindingor interaction molecules”. Binding between polypeptides of the inventionand such molecules, including receptor or binding or interactionmolecules may be exclusive to polypeptides of the invention, which isvery highly preferred, or it may be highly specific for polypeptides ofthe invention, which is highly preferred, or it may be highly specificto a group of proteins that includes polypeptides of the invention,which is preferred, or it may be specific to several groups of proteinsat least one of which includes polypeptides of the invention.

[0052] Receptors also may be non-naturally occurring, such as antibodiesand antibody-derived reagents that bind to polypeptides of theinvention.

DETAILED DESCRIPTION OF THE INVENTION

[0053] The present invention relates to novel lung specific polypeptidesand polynucleotides, referred to herein as LSGs, among other things, asdescribed in greater detail below.

[0054] Polynucleotides

[0055] In accordance with one aspect of the present invention, there areprovided isolated LSG polynucleotides which encode LSG polypeptides.

[0056] Using the information provided herein, such as the polynucleotidesequences set out in SEQ ID NO:1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 37 or 38 a polynucleotide of thepresent invention encoding a LSG may be obtained using standard cloningand screening procedures, such as those for cloning cDNAs using mRNAfrom cells of a human tumor as starting material.

[0057] Polynucleotides of the present invention may be in the form ofRNA, such as mRNA, or in the form of DNA, including, for instance, CDNAand genomic DNA obtained by cloning or produced by chemical synthetictechniques or by a combination thereof. The DNA may be double-strandedor single-stranded. Single-stranded DNA may be the coding strand, alsoknown as the sense strand, or it may be the non-coding strand, alsoreferred to as the anti-sense strand.

[0058] The coding sequence which encodes the polypeptides may beidentical to the coding sequence of the polynucleotides of SEQ ID NO:1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21,22, 37 or 38.

[0059] It also may be a polynucleotide with a different sequence, which,as a result of the redundancy (degeneracy) of the genetic code, encodesthe same polypeptides as encoded by SEQ ID NO:1, 2, 3, 4, 5, 6, 7, 8, 9,10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 37 or 38.

[0060] Polynucleotides of the present invention, such as SEQ ID NO: 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21,22, 37 or 38 which encode these polypeptides may comprise the codingsequence for the mature polypeptide by itself. Polynucleotides of thepresent invention may also comprise the coding sequence for the maturepolypeptide and additional coding sequences such as those encoding aleader or secretory sequence such as a pre-, or pro- or prepro-proteinsequence. Polynucleotides of the present invention may also comprise thecoding sequence of the mature polypeptide, with or without theaforementioned additional coding sequences, together with additional,non-coding sequences. Examples of additional non-coding sequences whichmay be incorporated into the polynucleotide of the present inventioninclude, but are not limited to, introns and non-coding 5′ and 3′sequences such as transcribed, non-translated sequences that play a rolein transcription, mRNA processing including, for example, splicing andpolyadenylation signals, ribosome binding and stability of mRNA, andadditional coding sequence which codes for amino acids such as thosewhich provide additional functionalities. Thus, for instance, thepolypeptide may be fused to a marker sequence such as a peptide whichfacilitates purification of the fused polypeptide. In certain preferredembodiments of this aspect of the invention, the marker sequence is ahexa-histidine peptide, such as the tag provided in the pQE vector(Qiagen, Inc.), among others, many of which are commercially available.As described in Gentz et al. (Proc. Natl. Acad. Sci., USA 86: 821-824(1989)), for instance, hexa-histidine provides for convenientpurification of the fusion protein. The HA tag corresponds to an epitopederived of influenza hemagglutinin protein (Wilson et al., Cell 37: 767(1984)).

[0061] In accordance with the foregoing, the term “polynucleotideencoding a polypeptide” as used herein encompasses polynucleotides whichinclude a sequence encoding a polypeptide of the present invention,particularly SEQ ID NO:1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14,15, 16, 17, 18, 19, 20, 21, 22, 37 or 38. Exemplary polypeptides encodedby the polynucleotides are depicted in SEQ ID NO: 39, 40, 41, 42, 43,44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, or 56. The termencompasses polynucleotides that include a single continuous region ordiscontinuous regions encoding the polypeptide (for example, interruptedby introns) together with additional regions, that also may containcoding and/or non-coding sequences.

[0062] The present invention further relates to variants of the hereinabove described polynucleotides which encode for fragments, analogs andderivatives of the LSG polypeptides. A variant of the polynucleotide maybe a naturally occurring variant such as a naturally occurring allelicvariant, or it may be a variant that is not known to occur naturally.Such non-naturally occurring variants of the polynucleotide may be madeby mutagenesis techniques, including those applied to polynucleotides,cells or organisms.

[0063] Among variants in this regard are variants that differ from theaforementioned polynucleotides by nucleotide substitutions, deletions oradditions. The substitutions, deletions or additions may involve one ormore nucleotides. The variants may be altered in coding or non-codingregions or both. Alterations in the coding regions may produceconservative or non-conservative amino acid substitutions, deletions oradditions.

[0064] Among the particularly preferred embodiments of the invention inthis regard are polynucleotides encoding polypeptides having the sameamino acid sequence encoded by a LSG polynucleotide comprising SEQ IDNO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19,20, 21, 22, 37 or 38; variants, analogs, derivatives and fragmentsthereof, and fragments of the variants, analogs and derivatives.Exemplary polypeptides encoded by these polynucleotides are depicted inSEQ ID NO:39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53,54, 55 or 56. Further particularly preferred in this regard are LSGpolynucleotides encoding polypeptide variants, analogs, derivatives andfragments, and variants, analogs and derivatives of the fragments, inwhich several, a few, 5 to 10, 1 to 5, 1 to 3, 2, 1 or no amino acidresidues are substituted, deleted or added, in any combination.Especially preferred among these are silent substitutions, additions anddeletions, which do not alter the properties and activities of the LSG.Also especially preferred in this regard are conservative substitutions.Most highly preferred are polynucleotides encoding polypeptides havingthe amino acid sequences as polypeptides encoded by SEQ ID NO: 1, 2, 3,4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 37or 38 without substitutions.

[0065] Further preferred embodiments of the invention are LSGpolynucleotides that are at least 70% identical to a polynucleotide ofSEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17,18, 19, 20, 21, 22, 37 or 38 and polynucleotides which are complementaryto such polynucleotides. More preferred are LSG polynucleotides thatcomprise a region that is at least 80% identical to a polynucleotide ofSEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17,18, 19, 20, 21, 22, 37 or 38. In this regard, LSG polynucleotides atleast 90% identical to the same are particularly preferred, and amongthese particularly preferred LSG polynucleotides, those with at least95% are especially preferred. Furthermore, those with at least 97% arehighly preferred among those with at least 95%, and among these thosewith at least 98% and at least 99% are particularly highly preferred,with at least 99% being the most preferred.

[0066] Particularly preferred embodiments in this respect, moreover, arepolynucleotides which encode polypeptides which retain substantially thesame biological function or activity as the mature polypeptides encodedby a polynucleotide of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 37 or 38.

[0067] The present invention further relates to polynucleotides thathybridize to the herein above-described LSG sequences. In this regard,the present invention especially relates to polynucleotides whichhybridize under stringent conditions to the herein above-describedpolynucleotides. As herein used, the term “stringent conditions” meanshybridization will occur only if there is at least 95% and preferably atleast 97% identity between the sequences.

[0068] As discussed additionally herein regarding polynucleotide assaysof the invention, for instance, polynucleotides of the invention asdescribed herein, may be used as a hybridization probe for cDNA andgenomic DNA to isolate full-length cDNAs and genomic clones encodingLSGs and to isolate cDNA and genomic clones of other genes that have ahigh sequence similarity to these LSGs. Such probes generally willcomprise at least 15 bases. Preferably, such probes will have at least30 bases and may have at least 50 bases.

[0069] For example, the coding region of LSG of the present inventionmay be isolated by screening using an oligonucleotide probe synthesizedfrom the known DNA sequence. A labeled oligonucleotide having a sequencecomplementary to that of a gene of the present invention is used toscreen a library of human cDNA, genomic DNA or mRNA to determine whichmembers of the library the probe hybridizes with.

[0070] The polynucleotides and polypeptides of the present invention maybe employed as research reagents and materials for discovery oftreatments and diagnostics to human disease, as further discussed hereinrelating to polynucleotide assays, inter alia.

[0071] The polynucleotides may encode a polypeptide which is the matureprotein plus additional amino or carboxyl-terminal amino acids, or aminoacids interior to the mature polypeptide (when the mature form has morethan one polypeptide chain, for instance). Such sequences may play arole in processing of a protein from precursor to a mature form, mayfacilitate/protein trafficking, may prolong or shorten protein half-lifeor may facilitate manipulation of a protein for assay or production,among other things. As generally is the case in situ, the additionalamino acids may be processed away from the mature protein by cellularenzymes.

[0072] A precursor protein having the mature form of the polypeptidefused to one or more prosequences may be an inactive form of thepolypeptide. When prosequences are removed, such inactive precursorsgenerally are activated. Some or all of the prosequences may be removedbefore activation. Generally, such precursors are called proproteins.

[0073] In sum, a polynucleotide of the present invention may encode amature protein, a mature protein plus a leader sequence (which may bereferred to as a preprotein), a precursor of a mature protein having oneor more prosequences which are not the leader sequences of a preprotein,or a preproprotein, which is a precursor to a proprotein, having aleader sequence and one or more prosequences, which generally areremoved during processing steps that produce active and mature forms ofthe polypeptide.

[0074] Polypeptides

[0075] The present invention further relates to LSG polypeptides,preferably polypeptides encoded by a polynucleotide of SEQ ID NO: 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22,37 or 38. Exemplary polypeptides are depicted in SEQ ID NO: 39, 40, 41,42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55 or 56. Theinvention also relates to fragments, analogs and derivatives of thesepolypeptides. The terms “fragment,” “derivative” and “analog” whenreferring to the polypeptides of the present invention means apolypeptide which retains essentially the same biological function oractivity as such polypeptides. Thus, an analog includes a proproteinwhich can be activated by cleavage of the proprotein portion to producean active mature polypeptide.

[0076] The polypeptide of the present invention may be a recombinantpolypeptide, a natural polypeptide or a synthetic polypeptide. Incertain preferred embodiments it is a recombinant polypeptide.

[0077] The fragment, derivative or analog of a polypeptide of or thepresent invention may be (I) one in which one or more of the amino acidresidues are substituted with a conserved or non-conserved amino acidresidue (preferably a conserved amino acid residue) and such substitutedamino acid residue may or may not be one encoded by the genetic code;(ii) one in which one or more of the amino acid residues includes asubstituent group; (iii) one in which the mature polypeptide is fusedwith another compound, such as a compound to increase the half-life ofthe polypeptide (for example, polyethylene glycol); or (iv) one in whichthe additional amino acids are fused to the mature polypeptide, such asa leader or secretory sequence or a sequence which is employed forpurification of the mature polypeptide or a proprotein sequence. Suchfragments, derivatives and analogs are deemed to be within the scope ofthose skilled in the art from the teachings herein. Among preferredvariants are those that vary from a reference by conservative amino acidsubstitutions. Such substitutions are those that substitute a givenamino acid in a polypeptide by another amino acid of likecharacteristics. Typically seen as conservative substitutions are thereplacements, one for another, among the aliphatic amino acids Ala, Val,Leu and Ile; interchange of the hydroxyl residues Ser and Thr, exchangeof the acidic residues Asp and Glu, substitution between the amideresidues Asn and Gln, exchange of the basic residues Lys and Arg andreplacements among the aromatic residues Phe, Tyr.

[0078] The polypeptides and polynucleotides of the present invention arepreferably provided in an isolated form, and preferably are purified tohomogeneity.

[0079] The polypeptides of the present invention include thepolypeptides encoded by the polynucleotide of SEQ ID NO: 1, 2, 3, 4, 5,6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 37 or 38(in particular the mature polypeptide) as well as polypeptides whichhave at least 75% similarity (preferably at least 75% identity), morepreferably at least 90% similarity (more preferably at least 90%identity), still more preferably at least 95% similarity (still morepreferably at least 95% identity), to a polypeptide encoded by SEQ IDNO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19,20, 21, 22, 37 or 38. Also included are portions of such polypeptidesgenerally containing at least 30 amino acids and more preferably atleast 50 amino acids. Exemplary polypeptides are depicted in SEQ IDNO:39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55 or56.

[0080] As known in the art “similarity” between two polypeptides isdetermined by comparing the amino acid sequence and its conserved aminoacid substitutes of one polypeptide sequence with that of a secondpolypeptide.

[0081] Fragments or portions of the polypeptides of the presentinvention may be employed for producing the corresponding full-lengthpolypeptide by peptide synthesis; therefore, the fragments may beemployed as intermediates for producing the full-length polypeptides.Fragments or portions of the polynucleotides of the present inventionmay be used to synthesize full-length polynucleotides of the presentinvention.

[0082] Fragments

[0083] Also among preferred embodiments of this aspect of the presentinvention are polypeptides comprising fragments of a polypeptide encodedby a polynucleotide of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 37 or 38. In this regard afragment is a polypeptide having an amino acid sequence that entirely isthe same as part but not all of the amino acid sequence of theaforementioned LSG polypeptides and variants or derivatives thereof.

[0084] Such fragments may be “free-standing,” i.e., not part of or fusedto other amino acids or polypeptides, or they may be contained within alarger polypeptide of which they form a part or region. When containedwithin a larger polypeptide, the presently discussed fragments mostpreferably form a single continuous region. However, several fragmentsmay be comprised within a single larger polypeptide. For instance,certain preferred embodiments relate to a fragment of a LSG polypeptideof the present comprised within a precursor polypeptide designed forexpression in a host and having heterologous pre- and pro-polypeptideregions fused to the amino terminus of the LSG fragment and anadditional region fused to the carboxyl terminus of the fragment.Therefore, fragments in one aspect of the meaning intended herein,refers to the portion or portions of a fusion polypeptide or fusionprotein derived from a LSG polypeptide.

[0085] As representative examples of polypeptide fragments of theinvention, there may be mentioned those which have from about 15 toabout 139 amino acids. In this context “about” includes the particularlyrecited range and ranges larger or smaller by several, a few, 5, 4, 3, 2or 1 amino acid at either extreme or at both extremes. Highly preferredin this regard are the recited ranges plus or minus as many as 5 aminoacids at either or at both extremes. Particularly highly preferred arethe recited ranges plus or minus as many as 3 amino acids at either orat both the recited extremes. Especially preferred are ranges plus orminus 1 amino acid at either or at both extremes or the recited rangeswith no additions or deletions. Most highly preferred of all in thisregard are fragments from about 15 to about 45 amino acids.

[0086] Among especially preferred fragments of the invention aretruncation mutants of the LSG polypeptides. Truncation mutants includeLSG polypeptides having an amino acid sequence encoded by apolynucleotide of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,14, 15, 16, 17, 18, 19, 20, 21, 22, 37 or 38 or variants or derivativesthereof, except for deletion of a continuous series of residues (thatis, a continuous region, part or portion) that includes the aminoterminus, or a continuous series of residues that includes the carboxylterminus or, as in double truncation mutants, deletion of two continuousseries of residues, one including the amino terminus and one includingthe carboxyl terminus. Fragments having the size ranges set out hereinalso are preferred embodiments of truncation fragments, which areespecially preferred among fragments generally.

[0087] Also preferred in this aspect of the invention are fragmentscharacterized by structural or functional attributes of the LSGpolypeptides of the present invention. Preferred embodiments of theinvention in this regard include fragments that comprise alpha-helix andalpha-helix forming regions (“alpha-regions”), beta-sheet andbeta-sheet-forming regions (“beta-regions”), turn and turn-formingregions (“turn-regions”), coil and coil-forming regions(“coil-regions”), hydrophilic regions, hydrophobic regions, alphaamphipathic regions, beta amphipathic regions, flexible regions,surface-forming regions and high antigenic index regions of the LSGpolypeptides of the present invention. Regions of the aforementionedtypes are identified routinely by analysis of the amino acid sequencesencoded by the polynucleotides of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 37 or 38. Preferredregions include Garnier-Robson alpha-regions, beta-regions, turn-regionsand coil-regions, Chou-Fasman alpha-regions, beta-regions andturn-regions, Kyte-Doolittle hydrophilic regions and hydrophilicregions, Eisenberg alpha and beta amphipathic regions, Karplus-Schulzflexible regions, Emini surface-forming regions and Jameson-Wolf highantigenic index regions. Among highly preferred fragments in this regardare those that comprise regions of LSGs that combine several structuralfeatures, such as several of the features set out above. In this regard,the regions defined by selected residues of a LSG polypeptide which allare characterized by amino acid compositions highly characteristic ofturn-regions, hydrophilic regions, flexible-regions, surface-formingregions, and high antigenic index-regions, are especially highlypreferred regions. Such regions may be comprised within a largerpolypeptide or may be by themselves a preferred fragment of the presentinvention, as discussed above. It will be appreciated that the term“about” as used in this paragraph has the meaning set out aboveregarding fragments in general.

[0088] Further preferred regions are those that mediate activities ofLSG polypeptides. Most highly preferred in this regard are fragmentsthat have a chemical, biological or other activity of a LSG polypeptide,including those with a similar activity or an improved activity, or witha decreased undesirable activity. Highly preferred in this regard arefragments that contain regions that are homologs in sequence, or inposition, or in both sequence and to active regions of relatedpolypeptides, and which include lung specific-binding proteins. Amongparticularly preferred fragments in these regards are truncationmutants, as discussed above.

[0089] It will be appreciated that the invention also relates topolynucleotides encoding the aforementioned fragments, polynucleotidesthat hybridize to polynucleotides encoding the fragments, particularlythose that hybridize under stringent conditions, and polynucleotidessuch as PCR primers for amplifying polynucleotides that encode thefragments. In these regards, preferred polynucleotides are those thatcorrespond to the preferred fragments, as discussed above.

[0090] Fusion Proteins

[0091] In one embodiment of the present invention, the LSG polypeptidesof the present invention are preferably fused to other proteins. Thesefusion proteins can be used for a variety of applications. For example,fusion of the present polypeptides to His-tag, HA-tag, protein A, IgGdomains, and maltose binding protein facilitates purification. (See alsoEP A 394,827; Traunecker, et al., Nature 331: 84-86 (1988)) Similarly,fusion to IgG-1, IgG-3, and albumin increases the halflife time in vivo.Nuclear localization signals fused to the polypeptides of the presentinvention can target the protein to a specific subcellular localization,while covalent heterodimer or homodimers can increase or decrease theactivity of a fusion protein. Fusion proteins can also create chimericmolecules having more than one function. Finally, fusion proteins canincrease solubility and/or stability of the fused protein compared tothe non-fused protein. All of these types of fusion proteins describedabove can be made in accordance with well known protocols.

[0092] For example, a LSG polypeptide can be fused to an IgG moleculevia the following protocol. Briefly, the human Fc portion of the IgGmolecule is PCR amplified using primers that span the 5′ and 3′ ends ofthe sequence. These primers also have convenient restriction enzymesites that facilitate cloning into an expression vector, preferably amammalian expression vector. For example, if pC4 (Accession No. 209646)is used, the human Fc portion can be ligated into the BamHI cloningsite. In this protocol, the 3′ BamHI site must be destroyed. Next, thevector containing the human Fc portion is re-restricted with BamHIthereby linearizing the vector, and a LSG polynucleotide of the presentinvention is ligated into this BamHI site. It is preferred that thepolynucleotide is cloned without a stop codon, otherwise a fusionprotein will not be produced.

[0093] If the naturally occurring signal sequence is used to produce thesecreted protein, pC4 does not need a second signal peptide.Alternatively, if the naturally occurring signal sequence is not used,the vector can be modified to include a heterologous signal sequence.(See, e. g., WO 96/34891.)

[0094] Diagnostic Assays

[0095] The present invention also relates to diagnostic assays andmethods, both quantitative and qualitative for detecting, diagnosing,monitoring, staging and prognosticating cancers by comparing levels ofLSG in a human patient with those of LSG in a normal human control. Forpurposes of the present invention, what is meant by LSG levels is, amongother things, native protein expressed by a gene comprising thepolynucleotide sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11,12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 37 or 38. Exemplarypolypeptides encoded by these polynucleotides are depicted in SEQ IDNO:39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55 or56. By “LSG” it is also meant herein polynucleotides which, due todegeneracy in genetic coding, comprise variations in nucleotide sequenceas compared to SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14,15, 16, 17, 18, 19, 20, 21, 22, 37 or 38 but which still encode the sameprotein. The native protein being detected may be whole, a breakdownproduct, a complex of molecules or chemically modified. In thealternative, what is meant by LSG as used herein, means the native mRNAencoded by a polynucleotide sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7,8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or 22, or a contigof SEQ ID NO:19 or 21, depicted as SEQ ID NO: 37 or 38, respectively,levels of the gene comprising the polynucleotide sequence of SEQ ID NO:1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,21, or 22, or levels of a polynucleotide which is capable of hybridizingunder stringent conditions to the antisense sequence of SEQ ID NO: 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22,37 or 38. Such levels are preferably determined in at least one ofcells, tissues and/or bodily fluids, including determination of normaland abnormal levels. Thus, for instance, a diagnostic assay inaccordance with the invention for diagnosing overexpression of LSGprotein compared to normal control bodily fluids, cells, or tissuesamples may be used to diagnose the presence of lung cancer.

[0096] All the methods of the present invention may optionally includedetermining the levels of other cancer markers as well as LSG. Othercancer markers, in addition to LSG, useful in the present invention willdepend on the cancer being tested and are known to those of skill in theart.

[0097] The present invention provides methods for diagnosing thepresence of lung cancer by analyzing for changes in levels of LSG incells, tissues or bodily fluids compared with levels of LSG in cells,tissues or bodily fluids of preferably the same type from a normal humancontrol, wherein an increase in levels of LSG in the patient versus thenormal human control is associated with the presence of lung cancer.

[0098] Without limiting the instant invention, typically, for aquantitative diagnostic assay a positive result indicating the patientbeing tested has cancer is one in which cells, tissues or bodily fluidlevels of the cancer marker, such as LSG, are at least two times higher,and most preferably are at least five times higher, than in preferablythe same cells, tissues or bodily fluid of a normal human control.

[0099] The present invention also provides a method of diagnosingmetastatic lung cancer in a patient having lung cancer which has not yetmetastasized for the onset of metastasis. In the method of the presentinvention, a human cancer patient suspected of having lung cancer whichmay have metastasized (but which was not previously known to havemetastasized) is identified. This is accomplished by a variety of meansknown to those of skill in the art.

[0100] In the present invention, determining the presence of LSG levelsin cells, tissues or bodily fluid, is particularly useful fordiscriminating between lung cancer which has not metastasized and lungcancer which has metastasized. Existing techniques have difficultydiscriminating between lung cancer which has metastasized and lungcancer which has not metastasized and proper treatment selection isoften dependent upon such knowledge.

[0101] In the present invention, the cancer marker levels measured insuch cells, tissues or bodily fluid is LSG, and are compared with levelsof LSG in preferably the same cells, tissue or bodily fluid type of anormal human control. That is, if the cancer marker being observed isjust LSG in serum, this level is preferably compared with the level ofLSG in serum of a normal human control. An increase in the LSG in thepatient versus the normal human control is associated with lung cancerwhich has metastasized.

[0102] Without limiting the instant invention, typically, for aquantitative diagnostic assay a positive result indicating the cancer inthe patient being tested or monitored has metastasized is one in whichcells, tissues or bodily fluid levels of the cancer marker, such as LSG,are at least two times higher, and most preferably are at least fivetimes higher, than in preferably the same cells, tissues or bodily fluidof a normal patient.

[0103] Normal human control as used herein includes a human patientwithout cancer and/or non cancerous samples from the patient; in themethods for diagnosing or monitoring for metastasis, normal humancontrol may preferably also include samples from a human patient that isdetermined by reliable methods to have lung cancer which has notmetastasized.

[0104] Staging

[0105] The invention also provides a method of staging lung cancer in ahuman patient. The method comprises identifying a human patient havingsuch cancer and analyzing cells, tissues or bodily fluid from such humanpatient for LSG. The LSG levels determined in the patient are thencompared with levels of LSG in preferably the same cells, tissues orbodily fluid type of a normal human control, wherein an increase in LSGlevels in the human patient versus the normal human control isassociated with a cancer which is progressing and a decrease in thelevels of LSG (but still increased over true normal levels) isassociated with a cancer which is regressing or in remission.

[0106] Monitoring

[0107] Further provided is a method of monitoring lung cancer in a humanpatient having such cancer for the onset of metastasis. The methodcomprises identifying a human patient having such cancer that is notknown to have metastasized; periodically analyzing cells, tissues orbodily fluid from such human patient for LSG; and comparing the LSGlevels determined in the human patient with levels of LSG in preferablythe same cells, tissues or bodily fluid type of a normal human control,wherein an increase in LSG levels in the human patient versus the normalhuman control is associated with a cancer which has metastasized. Inthis method, normal human control samples may also include prior patientsamples.

[0108] Further provided by this invention is a method of monitoring thechange in stage of lung cancer in a human patient having such cancer.The method comprises identifying a human patient having such cancer;periodically analyzing cells, tissues or bodily fluid from such humanpatient for LSG; and comparing the LSG levels determined in the humanpatient with levels of LSG in preferably the same cells, tissues orbodily fluid type of a normal human control, wherein an increase in LSGlevels in the human patient versus the normal human control isassociated with a cancer which is progressing in stage and a decrease inthe levels of LSG is associated with a cancer which is regressing instage or in remission. In this method, normal human control samples mayalso include prior patient samples.

[0109] Monitoring a patient for onset of metastasis is periodic andpreferably done on a quarterly basis. However, this may be done more orless frequently depending on the cancer, the particular patient, and thestage of the cancer.

[0110] Prognostic Testing and Clinical Trial Monitoring

[0111] The methods described herein can further be utilized asprognostic assays to identify subjects having or at risk of developing adisease or disorder associated with increased levels of LSG. The presentinvention provides a method in which a test sample is obtained from ahuman patient and LSG is detected. The presence of higher LSG levels ascompared to normal human controls is diagnostic for the human patientbeing at risk for developing cancer, particularly lung cancer.

[0112] The effectiveness of therapeutic agents to decrease expression oractivity of the LSGs of the invention can also be monitored by analyzinglevels of expression of the LSGs in a human patient in clinical trialsor in in vitro screening assays such as in human cells. In this way, thegene expression pattern can serve as a marker, indicative of thephysiological response of the human patient, or cells as the case maybe, to the agent being tested.

[0113] Detection of Genetic Lesions or Mutations

[0114] The methods of the present invention can also be used to detectgenetic lesions or mutations in LSG, thereby determining if a human withthe genetic lesion is at risk for lung cancer or has lung cancer.Genetic lesions can be detected, for example, by ascertaining theexistence of a deletion and/or addition and/or substitution of one ormore nucleotides from the LSGs of this invention, a chromosomalrearrangement of LSG, aberrant modification of LSG (such as of themethylation pattern of the genomic DNA), the presence of a non-wild typesplicing pattern of a mRNA transcript of LSG, allelic loss of LSG,and/or inappropriate post-translational modification of LSG protein.Methods to detect such lesions in the LSG of this invention are known tothose of skill in the art.

[0115] For example, in one embodiment, alterations in a genecorresponding to a LSG polynucleotide of the present invention aredetermined via isolation of RNA from entire families or individualpatients presenting with a phenotype of interest (such as a disease) isbe isolated. cDNA is then generated from these RNA samples usingprotocols known in the art. See, e.g. Sambrook et al. (MOLECULARCLONING: A LABORATORY MANUAL, 2nd Ed., Cold Spring Harbor LaboratoryPress, Cold Spring Harbor, N.Y. (1989)), which is illustrative of themany laboratory manuals that detail these techniques. The cDNA is thenused as a template for PCR, employing primers surrounding regions ofinterest in SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14,15, 16, 17, 18, 19, 20, 21, 22, 37 or 38. PCR conditions typicallyconsist of 35 cycles at 95° C. for 30 seconds; 60-120 seconds at 52-58°C.; and 60-120 seconds at 70° C., using buffer solutions described inSidransky, D., et al., Science 252: 706 (1991). PCR products aresequenced using primers labeled at their 5′ end with T4 polynucleotidekinase, employing SequiTherm Polymerase (Epicentre Technologies). Theintron-exon borders of selected exons are also determined and genomicPCR products analyzed to confirm the results. PCR products harboringsuspected mutations are then cloned and sequenced to validate theresults of the direct sequencing. PCR products are cloned into T-tailedvectors as described in Holton, T. A. and Graham, M. W., Nucleic AcidsResearch, 19 : 1156 (1991) and sequenced with T7 polymerase (UnitedStates Biochemical). Affected individuals are identified by mutationsnot present in unaffected individuals.

[0116] Genomic rearrangements can also be observed as a method ofdetermining alterations in a gene corresponding to a polynucleotide. Inthis method, genomic clones are nick-translated with digoxigenindeoxy-uridine 5′ triphosphate (Boehringer Manheim), and FISH isperformed as described in Johnson, C. et al., Methods Cell Biol. 35:73-99 (1991). Hybridization with a labeled probe is carried out using avast excess of human DNA for specific hybridization to the correspondinggenomic locus. Chromosomes are counterstained with4,6-diamino-2-phenylidole and propidium iodide, producing a combinationof C-and R-bands. Aligned images for precise mapping are obtained usinga triple-band filter set (Chroma Technology, Brattleboro, Vt.) incombination with a cooled charge-coupled device camera (Photometrics,Tucson, Ariz.) and variable excitation wavelength filters (Johnson etal., Genet. Anal. Tech. Appl., 8: 75 (1991)). Image collection, analysisand chromosomal fractional length measurements are performed using theISee Graphical Program System (Inovision Corporation, Durham, N.C.).Chromosome alterations of the genomic region hybridized by the probe areidentified as insertions, deletions, and translocations. Thesealterations are used as a diagnostic marker for an associated disease.

[0117] Assay Techniques

[0118] Assay techniques that can be used to determine levels of geneexpression (including protein levels), such as LSG of the presentinvention, in a sample derived from a patient are well known to those ofskill in the art. Such assay methods include, without limitation,radioimmunoassays, reverse transcriptase PCR (RT-PCR) assays,immunohistochemistry assays, in situ hybridization assays,competitive-binding assays, Western Blot analyses, ELISA assays andproteomic approaches: two-dimensional gel electrophoresis (2Delectrophoresis) and non-gel based approaches such as mass spectrometryor protein interaction profiling. Among these, ELISAs are frequentlypreferred to diagnose a gene's expressed protein in biological fluids.

[0119] An ELISA assay initially comprises preparing an antibody, if notreadily available from a commercial source, specific to LSG, preferablya monoclonal antibody. In addition a reporter antibody generally isprepared which binds specifically to LSG. The reporter antibody isattached to a detectable reagent such as radioactive, fluorescent orenzymatic reagent, for example horseradish peroxidase enzyme or alkalinephosphatase.

[0120] To carry out the ELISA, antibody specific to LSG is incubated ona solid support, e.g. a polystyrene dish, that binds the antibody. Anyfree protein binding sites on the dish are then covered by incubatingwith a non-specific protein such as bovine serum albumin. Next, thesample to be analyzed is incubated in the dish, during which time LSGbinds to the specific antibody attached to the polystyrene dish. Unboundsample is washed out with buffer. A reporter antibody specificallydirected to LSG and linked to a detectable reagent such as horseradishperoxidase is placed in the dish resulting in binding of the reporterantibody to any monoclonal antibody bound to LSG. Unattached reporterantibody is then washed out. Reagents for peroxidase activity, includinga calorimetric substrate are then added to the dish. Immobilizedperoxidase, linked to LSG antibodies, produces a colored reactionproduct. The amount of color developed in a given time period isproportional to the amount of LSG protein present in the sample.Quantitative results typically are obtained by reference to a standardcurve.

[0121] A competition assay can also be employed wherein antibodiesspecific to LSG are attached to a solid support and labeled LSG and asample derived from the host are passed over the solid support. Theamount of label detected which is attached to the solid support can becorrelated to a quantity of LSG in the sample.

[0122] Using all or a portion of a nucleic acid sequence of LSG of thepresent invention as a hybridization probe, nucleic acid methods canalso be used to detect LSG mRNA as a marker for lung cancer. Polymerasechain reaction (PCR) and other nucleic acid methods, such as ligasechain reaction (LCR) and nucleic acid sequence based amplification(NASBA), can be used to detect malignant cells for diagnosis andmonitoring of various malignancies. For example, reverse-transcriptasePCR (RT-PCR) is a powerful technique which can be used to detect thepresence of a specific mRNA population in a complex mixture of thousandsof other mRNA species. In RT-PCR, an mRNA species is first reversetranscribed to complementary DNA (cDNA) with use of the enzyme reversetranscriptase; the cDNA is then amplified as in a standard PCR reaction.RT-PCR can thus reveal by amplification the presence of a single speciesof mRNA. Accordingly, if the mRNA is highly specific for the cell thatproduces it, RT-PCR can be used to identify the presence of a specifictype of cell.

[0123] Hybridization to clones or oligonucleotides arrayed on a solidsupport (i.e. gridding) can be used to both detect the expression of andquantitate the level of expression of that gene. In this approach, acDNA encoding the LSG gene is fixed to a substrate. The substrate may beof any suitable type including but not limited to glass, nitrocellulose,nylon or plastic. At least a portion of the DNA encoding the LSG gene isattached to the substrate and then incubated with the analyte, which maybe RNA or a complementary DNA (cDNA) copy of the RNA, isolated from thetissue of interest. Hybridization between the substrate bound DNA andthe analyte can be detected and quantitated by several means includingbut not limited to radioactive labeling or fluorescence labeling of theanalyte or a secondary molecule designed to detect the hybrid.Quantitation of the level of gene expression can be done by comparisonof the intensity of the signal from the analyte compared with thatdetermined from known standards. The standards can be obtained by invitro transcription of the target gene, quantitating the yield, and thenusing that material to generate a standard curve.

[0124] Of the proteomic approaches, 2D electrophoresis is a techniquewell known to those in the art. Isolation of individual proteins from asample such as serum is accomplished using sequential separation ofproteins by different characteristics usually on polyacrylamide gels.First, proteins are separated by size using an electric current. Thecurrent acts uniformly on all proteins, so smaller proteins move fartheron the gel than larger proteins. The second dimension applies a currentperpendicular to the first and separates proteins not on the basis ofsize but on the specific electric charge carried by each protein. Sinceno two proteins with different sequences are identical on the basis ofboth size and charge, the result of a 2D separation is a square gel inwhich each protein occupies a unique spot. Analysis of the spots withchemical or antibody probes, or subsequent protein microsequencing canreveal the relative abundance of a given protein and the identity of theproteins in the sample.

[0125] The above tests can be carried out on samples derived from avariety of cells, bodily fluids and/or tissue extracts such ashomogenates or solubilized tissue obtained from a patient. Tissueextracts are obtained routinely from tissue biopsy and autopsy material.Bodily fluids useful in the present invention include blood, urine,saliva or any other bodily secretion or derivative thereof. By blood itis meant to include whole blood, plasma, serum or any derivative ofblood.

[0126] In Vivo Targeting of LSG/Lung Cancer Therapy

[0127] Identification of this LSG is also useful in the rational designof new therapeutics for imaging and treating cancers, and in particularlung cancer. For example, in one embodiment, antibodies whichspecifically bind to LSG can be raised and used in vivo in patientssuspected of suffering from lung cancer. Antibodies which specificallybind LSG can be injected into a patient suspected of having lung cancerfor diagnostic and/or therapeutic purposes. Thus, another aspect of thepresent invention provides for a method for preventing the onset andtreatment of lung cancer in a human patient in need of such treatment byadministering to the patient an effective amount of antibody. By“effective amount” it is meant the amount or concentration of antibodyneeded to bind to the target antigens expressed on the tumor to causetumor shrinkage for surgical removal, or disappearance of the tumor. Thebinding of the antibody to the overexpressed LSG is believed to causethe death of the cancer cell expressing such LSG. The preparation anduse of antibodies for in vivo diagnosis and treatment is well known inthe art. For example, antibody-chelators labeled with Indium-111 havebeen described for use in the radioimmunoscintographic imaging ofcarcinoembryonic antigen expressing tumors (Sumerdon et al. Nucl. Med.Biol. 1990 17:247-254). In particular, these antibody-chelators havebeen used in detecting tumors in patients suspected of having recurrentcolorectal cancer (Griffin et al. J. Clin. Onc. 1991 9:631-640).Antibodies with paramagnetic ions as labels for use in magneticresonance imaging have also been described (Lauffer, R. B. MagneticResonance in Medicine 1991 22:339-342). Antibodies directed against LSGcan be used in a similar manner. Labeled antibodies which specificallybind LSG can be injected into patients suspected of having lung cancerfor the purpose of diagnosing or staging of the disease status of thepatient. The label used will be selected in accordance with the imagingmodality to be used. For example, radioactive labels such as Indium-111,Technetium-99m or Iodine-131 can be used for planar scans or singlephoton emission computed tomography (SPECT). Positron emitting labelssuch as Fluorine-19 can be used in positron emission tomography.Paramagnetic ions such as Gadlinium (III) or Manganese (II) can be usedin magnetic resonance imaging (MRI). Presence of the label, as comparedto imaging of normal tissue, permits determination of the spread of thecancer. The amount of label within an organ or tissue also allowsdetermination of the presence or absence of cancer in that organ ortissue.

[0128] Antibodies which can be used in in vivo methods includepolyclonal, monoclonal and omniclonal antibodies and antibodies preparedvia molecular biology techniques. Antibody fragments and aptamers andsingle-stranded oligonucleotides such as those derived from an in vitroevolution protocol referred to as SELEX and well known to those skilledin the art can also be used.

[0129] Screening Assays

[0130] The present invention also provides methods for identifyingmodulators which bind to LSG protein or have a modulatory effect on theexpression or activity of LSG protein. Modulators which decrease theexpression or activity of LSG protein are believed to be useful intreating lung cancer. Such screening assays are known to those of skillin the art and include, without limitation, cell-based assays and cellfree assays.

[0131] Small molecules predicted via computer imaging to specificallybind to regions of LSG can also be designed, synthesized and tested foruse in the imaging and treatment of lung cancer. Further, libraries ofmolecules can be screened for potential anticancer agents by assessingthe ability of the molecule to bind to the LSGs identified herein.Molecules identified in the library as being capable of binding to LSGare key candidates for further evaluation for use in the treatment oflung cancer. In a preferred embodiment, these molecules willdownregulate expression and/or activity of LSG in cells.

[0132] Adoptive Immunotherapy and Vaccines

[0133] Adoptive immunotherapy of cancer refers to a therapeutic approachin which immune cells with an antitumor reactivity are administered to atumor-bearing host, with the aim that the cells mediate either directlyor indirectly, the regression of an established tumor. Transfusion oflymphocytes, particularly T lymphocytes, falls into this category andinvestigators at the National Cancer Institute (NCI) have usedautologous reinfusion of peripheral blood lymphocytes ortumor-infiltrating lymphocytes (TIL), T cell cultures from biopsies ofsubcutaneous lymph nodules, to treat several human cancers (Rosenberg,S. A., U.S. Pat. No. 4,690,914, issued Sep. 1, 1987; Rosenberg, S. A.,et al., 1988, N. England J. Med. 319:1676-1680).

[0134] The present invention relates to compositions and methods ofadoptive immunotherapy for the prevention and/or treatment of primaryand metastatic lung cancer in humans using macrophages sensitized to theantigenic LSG molecules, with or without non-covalent complexes of heatshock protein (hsp). Antigenicity or immunogenicity of the LSG isreadily confirmed by the ability of the LSG protein or a fragmentthereof to raise antibodies or educate naive effector cells, which inturn lyse target cells expressing the antigen (or epitope).

[0135] Cancer cells are, by definition, abnormal and contain proteinswhich should be recognized by the immune system as foreign since theyare not present in normal tissues. However, the immune system oftenseems to ignore this abnormality and fails to attack tumors. The foreignLSG proteins that are produced by the cancer cells can be used to revealtheir presence. The LSG is broken into short fragments, called tumorantigens, which are displayed on the surface of the cell. These tumorantigens are held or presented on the cell surface by molecules calledMHC, of which there are two types: class I and II. Tumor antigens inassociation with MHC class I molecules are recognized by cytotoxic Tcells while antigen-MHC class II complexes are recognized by a secondsubset of T cells called helper cells. These cells secrete cytokineswhich slow or stop tumor growth and help another type of white bloodcell, B cells, to make antibodies against the tumor cells.

[0136] In adoptive immunotherapy, T cells or other antigen presentingcells (APCs) are stimulated outside the body (ex vivo), using the tumorspecific LSG antigen. The stimulated cells are then reinfused into thepatient where they attack the cancerous cells. Research has shown thatusing both cytotoxic and helper T cells is far more effective than usingeither subset alone. Additionally, the LSG antigen may be complexed withheat shock proteins to stimulate the APCs as described in U.S. Pat. No.5,985,270.

[0137] The APCs can be selected from among those antigen presentingcells known in the art including, but not limited to, macrophages,dendritic cells, B lymphocytes, and a combination thereof, and arepreferably macrophages. In a preferred use, wherein cells are autologousto the individual, autologous immune cells such as lymphocytes,macrophages or other APCs are used to circumvent the issue of whom toselect as the donor of the immune cells for adoptive transfer. Anotherproblem circumvented by use of autologous immune cells is graft versushost disease which can be fatal if unsuccessfully treated.

[0138] In adoptive immunotherapy with gene therapy, DNA of the LSG canbe introduced into effector cells similarly as in conventional genetherapy. This can enhance the cytotoxicity of the effector cells totumor cells as they have been manipulated to produce the antigenicprotein resulting in improvement of the adoptive immunotherapy.

[0139] LSG antigens of this invention are also useful as components oflung cancer vaccines. The vaccine comprises an immunogenicallystimulatory amount of a LSG antigen. Immunogenically stimulatory amountrefers to that amount of antigen that is able to invoke the desiredimmune response in the recipient for the amelioration, or treatment oflung cancer. Effective amounts may be determined empirically by standardprocedures well known to those skilled in the art.

[0140] The LSG antigen may be provided in any one of a number of vaccineformulations which are designed to induce the desired type of immuneresponse, e.g., antibody and/or cell mediated. Such formulations areknown in the art and include, but are not limited to, formulations suchas those described in U.S. Pat. No. 5,585,103. Vaccine formulations ofthe present invention used to stimulate immune responses can alsoinclude pharmaceutically acceptable adjuvants.

[0141] Vectors, Host Cells, Expression

[0142] The present invention also relates to vectors which includepolynucleotides of the present invention, host cells which aregenetically engineered with vectors of the invention and the productionof polypeptides of the invention by recombinant techniques.

[0143] Host cells can be genetically engineered to incorporate LSGpolynucleotides and express LSG polypeptides of the present invention.For instance, LSG polynucleotides may be introduced into host cellsusing well known techniques of infection, transduction, transfection,transvection and transformation. The LSG polynucleotides may beintroduced alone or with other polynucleotides. Such otherpolynucleotides may be introduced independently, co-introduced orintroduced joined to the LSG polynucleotides of the invention.

[0144] For example, LSG polynucleotides of the invention may betransfected into host cells with another, separate, polynucleotideencoding a selectable marker, using standard techniques forco-transfection and selection in, for instance, mammalian cells. In thiscase, the polynucleotides generally will be stably incorporated into thehost cell genome.

[0145] Alternatively, the LSG polynucleotide may be joined to a vectorcontaining a selectable marker for propagation in a host. The vectorconstruct may be introduced into host cells by the aforementionedtechniques. Generally, a plasmid vector is introduced as DNA in aprecipitate, such as a calcium phosphate precipitate, or in a complexwith a charged lipid. Electroporation also may be used to introduce LSGpolynucleotides into a host. If the vector is a virus, it may bepackaged in vitro or introduced into a packaging cell and the packagedvirus may be transduced into cells. A wide variety of well knowntechniques conducted routinely by those of skill in the art are suitablefor making LSG polynucleotides and for introducing LSG polynucleotidesinto cells in accordance with this aspect of the invention. Suchtechniques are reviewed at length in reference texts such as Sambrook etal., previously cited herein.

[0146] Vectors which may be used in the present invention include, forexample, plasmid vectors, single- or double-stranded phage vectors, andsingle- or double-stranded RNA or DNA viral vectors. Such vectors may beintroduced into cells as polynucleotides, preferably DNA, by well knowntechniques for introducing DNA and RNA into cells. The vectors, in thecase of phage and viral vectors, also may be and preferably areintroduced into cells as packaged or encapsidated virus by well knowntechniques for infection and transduction. Viral vectors may bereplication competent or replication defective. In the latter case viralpropagation generally will occur only in complementing host cells.

[0147] Preferred vectors for expression of polynucleotides andpolypeptides of the present invention include, but are not limited to,vectors comprising cis-acting control regions effective for expressionin a host operatively linked to the polynucleotide to be expressed.Appropriate trans-acting factors either are supplied by the host,supplied by a complementing vector or supplied by the vector itself uponintroduction into the host.

[0148] In certain preferred embodiments in this regard, the vectorsprovide for specific expression. Such specific expression may beinducible expression or expression only in certain types of cells orboth inducible and cell-specific. Particularly preferred among induciblevectors are vectors that can be induced to express by environmentalfactors that are easy to manipulate, such as temperature and nutrientadditives. A variety of vectors suitable to this aspect of theinvention, including constitutive and inducible expression vectors foruse in prokaryotic and eukaryotic hosts, are well known and employedroutinely by those of skill in the art.

[0149] The engineered host cells can be cultured in conventionalnutrient media which may be modified as appropriate for, inter alia,activating promoters, selecting transformants or amplifying genes.Culture conditions such as temperature, pH and the like, previously usedwith the host cell selected for expression, generally will be suitablefor expression of LSG polypeptides of the present invention.

[0150] A great variety of expression vectors can be used to express LSGpolypeptides of the invention. Such vectors include chromosomal,episomal and virus-derived vectors. Vectors may be derived frombacterial plasmids, from bacteriophage, from yeast episomes, from yeastchromosomal elements, from viruses such as baculoviruses, papovaviruses, such as SV40, vaccinia viruses, adenoviruses, fowl pox viruses,pseudorabies viruses and retroviruses, and from combinations thereofsuch as those derived from plasmid and bacteriophage genetic elements,such cosmids and phagemids. All may be used for expression in accordancewith this aspect of the present invention. Generally, any vectorsuitable to maintain, propagate or express polynucleotides to express apolypeptide in a host may be used for expression in this regard.

[0151] The appropriate DNA sequence may be inserted into the vector byany of a variety of well-known and routine techniques. In general, a DNAsequence for expression is joined to an expression vector by cleavingthe DNA sequence and the expression vector with one or more restrictionendonucleases and then joining the restriction fragments together usingT4 DNA ligase. Procedures for restriction and ligation that can be usedto this end are well known and routine to those of skill. Suitableprocedures in this regard, and for constructing expression vectors usingalternative techniques, which also are well known and routine to thoseskill, are set forth in great detail in Sambrook et al. cited elsewhereherein.

[0152] The DNA sequence in the expression vector is operatively linkedto appropriate expression control sequence(s), including, for instance,a promoter to direct mRNA transcription. Representative promotersinclude the phage lambda PL promoter, the E. coli lac, trp and tacpromoters, the SV40 early and late promoters, and promoters ofretroviral LTRs, to name just a few of the well-known promoters. It willbe understood that numerous promoters not mentioned are also suitablefor use in this aspect of the invention and are well known and readilymay be employed by those of skill in the manner illustrated by thediscussion and the examples herein.

[0153] In general, expression constructs will contain sites fortranscription initiation and termination, and, in the transcribedregion, a ribosome binding site for translation. The coding portion ofthe mature transcripts expressed by the constructs will include atranslation initiating AUG at the beginning and a termination codonappropriately positioned at the end of the polypeptide to be translated.

[0154] In addition, the constructs may contain control regions thatregulate as well as engender expression. Generally, in accordance withmany commonly practiced procedures, such regions will operate bycontrolling transcription, such as repressor binding sites andenhancers, among others.

[0155] Vectors for propagation and expression generally will includeselectable markers. Such markers also may be suitable for amplificationor the vectors may contain additional markers for this purpose. In thisregard, the expression vectors preferably contain one or more selectablemarker genes to provide a phenotypic trait for selection of transformedhost cells. Preferred markers include dihydrofolate reductase orneomycin resistance for eukaryotic cell culture, and tetracycline orampicillin resistance genes for culturing in E. coli and other bacteria.

[0156] The vector containing the appropriate DNA sequence as describedelsewhere herein, as well as an appropriate promoter, and otherappropriate control sequences, may be introduced into an appropriatehost using a variety of well known techniques suitable to expressiontherein of a desired polypeptide. Representative examples of appropriatehosts include bacterial cells, such as E. coli, Streptomyces andSalmonella typhimurium cells; fungal cells, such as yeast cells; insectcells such as Drosophila S2 and Spodoptera Sf9 cells; animal cells suchas CHO, COS and Bowes melanoma cells; and plant cells. Hosts for a greatvariety of expression constructs are well known, and those of skill willbe enabled by the present disclosure readily to select a host forexpressing a LSG polypeptide in accordance with this aspect of thepresent invention.

[0157] More particularly, the present invention also includesrecombinant constructs, such as expression constructs, comprising one ormore of the sequences described above. The constructs comprise a vector,such as a plasmid or viral vector, into which such LSG sequence of theinvention has been inserted. The sequence may be inserted in a forwardor reverse orientation. In certain preferred embodiments in this regard,the construct further comprises regulatory sequences, including, forexample, a promoter, operably linked to the sequence. Large numbers ofsuitable vectors and promoters are known to those of skill in the art,and there are many commercially available vectors suitable for use inthe present invention.

[0158] The following vectors, which are commercially available, areprovided by way of example. Among vectors preferred for use in bacteriaare pQE70, pQE60 and pQE-9, available from Qiagen; pBS vectors,Phagescript vectors, Bluescript vectors, pNH8A, pNH16a, pNH18A, pNH46A,available from Stratagene; and ptrc99a, pKK223-3, pKK233-3, pDR540,pRIT5 available from Pharmacia. Among preferred eukaryotic vectors arePWLNEO, pSV2CAT, pOG44, pXT1 and pSG available from Stratagene; andpSVK3, PBPV, PMSG and pSVL available from Pharmacia. These vectors arelisted solely by way of illustration of the many commercially availableand well known vectors that are available to those of skill in the artfor use in accordance with this aspect of the present invention. It willbe appreciated by those of skill in the art upon reading this disclosurethat any other plasmid or vector suitable for introduction, maintenance,propagation and/or expression of a LSG polynucleotide or polypeptide ofthe invention in a host may be used in this aspect of the invention.

[0159] Promoter regions can be selected from any desired gene usingvectors that contain a reporter transcription unit lacking a promoterregion, such as a chloramphenicol acetyl transferase (“cat”)transcription unit, downstream of a restriction site or sites forintroducing a candidate promoter fragment; i.e., a fragment that maycontain a promoter. As is well known, introduction into the vector of apromoter-containing fragment at the restriction site upstream of the catgene engenders production of CAT activity detectable by standard CATassays. Vectors suitable to this end are well known and readilyavailable. Two such vectors are pKK232-8 and pCM7. Thus, promoters forexpression of LSG polynucleotides of the present invention include, notonly well known and readily available promoters, but also promoters thatreadily may be obtained by the foregoing technique, using a reportergene.

[0160] Among known bacterial promoters suitable for expression ofpolynucleotides and polypeptides in accordance with the presentinvention are the E. coli laci and lacZ promoters, the T3 and T7promoters, the gpt promoter, the lambda PR, PL promoters and the trppromoter.

[0161] Among known eukaryotic promoters suitable in this regard are theCMV immediate early promoter, the HSV thymidine kinase promoter, theearly and late SV40 promoters, the promoters of retroviral LTRs, such asthose of the Rous sarcoma virus (“RSV”), and metallothionein promoters,such as the mouse metallothionein-I promoter.

[0162] Selection of appropriate vectors and promoters for expression ina host cell is a well known procedure and the requisite techniques forexpression vector construction, introduction of the vector into the hostand expression in the host are routine skills in the art.

[0163] The present invention also relates to host cells containing theabove-described constructs. The host cell can be a higher eukaryoticcell, such as a mammalian cell, or a lower eukaryotic cell, such as ayeast cell. Alternatively, the host cell can be a prokaryotic cell, suchas a bacterial cell.

[0164] Introduction of the construct into the host cell can be effectedby calcium phosphate transfection, DEAE-dextran mediated transfection,cationic lipid-mediated transfection, electroporation, transduction,infection or other methods. Such methods are described in many standardlaboratory manuals, such as Davis et al. BASIC METHODS IN MOLECULARBIOLOGY, (1986).

[0165] Constructs in host cells can be used in a conventional manner toproduce the gene product encoded by the recombinant sequence.Alternatively, LSG polypeptides of the invention can be syntheticallyproduced by conventional peptide synthesizers.

[0166] Mature proteins can be expressed in mammalian cells, yeast,bacteria, or other cells under the control of appropriate promoters.Cell-free translation systems can also be employed to produce suchproteins using RNAs derived from the DNA constructs of the presentinvention. Appropriate cloning and expression vectors for use withprokaryotic and eukaryotic hosts are described by Sambrook et al. citedelsewhere herein.

[0167] Generally, recombinant expression vectors will include origins ofreplication, a promoter derived from a highly-expressed gene to directtranscription of a downstream structural sequence, and a selectablemarker to permit isolation of vector containing cells after exposure tothe vector. Among suitable promoters are those derived from the genesthat encode glycolytic enzymes such as 3-phosphoglycerate kinase(“PGK”), a-factor, acid phosphatase, and heat shock proteins, amongothers. Selectable markers include the ampicillin resistance gene of E.coli and the trpl gene of S. cerevisiae.

[0168] Transcription of DNA encoding the LSG polypeptides of the presentinvention by higher eukaryotes may be increased by inserting an enhancersequence into the vector. Enhancers are cis-acting elements of DNA,usually about from 10 to 300 base pairs (bp) that act to increasetranscriptional activity of a promoter in a given host cell-type.Examples of enhancers include the SV40 enhancer, which is located on thelate side of the replication origin at bp 100 to 270, thecytomegalovirus early promoter enhancer, the polyoma enhancer on thelate side of the replication origin, and adenovirus enhancers.

[0169] A polynucleotide of the present invention, encoding aheterologous structural sequence of a LSG polypeptide of the presentinvention, generally will be inserted into the vector using standardtechniques so that it is operably linked to the promoter for expression.The polynucleotide will be positioned so that the transcription startsite is located appropriately 5′ to a ribosome binding site. Theribosome binding site will be 5′ to the AUG that initiates translationof the polypeptide to be expressed. Generally, there will be no otheropen reading frames that begin with an initiation codon, usually AUG,lying between the ribosome binding site and the initiating AUG. Also,generally, there will be a translation stop codon at the end of thepolypeptide and there will be a polyadenylation signal and atranscription termination signal appropriately disposed at the 3′ end ofthe transcribed region.

[0170] Appropriate secretion signals may be incorporated into theexpressed polypeptide for secretion of the translated protein into thelumen of the endoplasmic reticulum, into the periplasmic space or intothe extracellular environment. The signals may be endogenous to thepolypeptide or they may be heterologous signals.

[0171] The polypeptide may be expressed in a modified form, such as afusion protein, and may include not only secretion signals but alsoadditional heterologous functional regions. Thus, for instance, a regionof additional amino acids, particularly charged amino acids, may beadded to the N-terminus of the polypeptide to improve stability andpersistence in the host cell during purification or during subsequenthandling and storage. A region also may be added to the polypeptide tofacilitate purification. Such regions may be removed prior to finalpreparation of the polypeptide. The addition of peptide moieties topolypeptides to engender secretion or excretion, to improve stabilityand to facilitate purification, among others, are familiar and routinetechniques in the art.

[0172] Suitable prokaryotic hosts for propagation, maintenance orexpression of LSG polynucleotides and polypeptides in accordance withthe invention include Escherichia coli, Bacillus subtilis and Salmonellatyphimurium. Various species of Pseudomonas, Streptomyces, andStaphylococcus are suitable hosts in this regard. Many other hosts alsoknown to those of skill may also be employed in this regard.

[0173] As a representative, but non-limiting example, useful expressionvectors for bacterial use can comprise a selectable marker and bacterialorigin of replication derived from commercially available plasmidscomprising genetic elements of the well known cloning vector pBR322.Such commercial vectors include, for example, pKK223-3 (Pharmacia FineChemicals, Uppsala, Sweden) and GEM1 (Promega Biotec, Madison, Wis.,USA). These pBR322 “backbone” sections are combined with an appropriatepromoter and the structural sequence to be expressed. Followingtransformation of a suitable host strain and growth of the host strainto an appropriate cell density, where the selected promoter is inducibleit is induced by appropriate means (e.g., temperature shift or exposureto chemical inducer) and cells are cultured for an additional period.Cells typically then are harvested by centrifugation, disrupted byphysical or chemical means, and the resulting crude extract retained forfurther purification. Microbial cells employed in expression of proteinscan be disrupted by any convenient method, including freeze-thawcycling, sonication, mechanical disruption, or use of cell lysingagents, such methods are well know to those skilled in the art.

[0174] Various mammalian cell culture systems can be employed forexpression, as well. An exemplary mammalian expression systems is theCOS-7 line of monkey kidney fibroblasts described in Gluzman et al.,Cell 23: 175 (1981). Other mammalian cell lines capable of expressing acompatible vector include for example, the C127, 3T3, CHO, HeLa, humankidney 293 and BHK cell lines. Mammalian expression vectors comprise anorigin of replication, a suitable promoter and enhancer, and anyribosome binding sites, polyadenylation sites, splice donor and acceptorsites, transcriptional termination sequences, and 5′ flankingnon-transcribed sequences that are necessary for expression. In certainpreferred embodiments in this regard DNA sequences derived from the SV40splice sites, and the SV40 polyadenylation sites are used for requirednon-transcribed genetic elements of these types.

[0175] LSG polypeptides can be recovered and purified from recombinantcell cultures by well-known methods including ammonium sulfate orethanol precipitation, acid extraction, anion or cation exchangechromatography, phosphocellulose chromatography, hydrophobic interactionchromatography, affinity chromatography, hydroxylapatite chromatographyand lectin chromatography. Most preferably, high performance liquidchromatography (“HPLC”) is employed for purification. Well knowntechniques for refolding proteins may be employed to regenerate activeconformation when the polypeptide is denatured during isolation and orpurification.

[0176] LSG polypeptides of the present invention include naturallypurified products, products of chemical synthetic procedures, andproducts produced by recombinant techniques from a prokaryotic oreukaryotic host, including, for example, bacterial, yeast, higher plant,insect and mammalian cells. Depending upon the host employed in arecombinant production procedure, the LSG polypeptides of the presentinvention may be glycosylated or may be non-glycosylated. In addition,LSG polypeptides of the invention may also include an initial modifiedmethionine residue, in some cases as a result of host-mediatedprocesses.

[0177] LSG polynucleotides and polypeptides may be used in accordancewith the present invention for a variety of applications, particularlythose that make use of the chemical and biological properties of theLSGs. Additional applications relate to diagnosis and to treatment ofdisorders of cells, tissues and organisms. These aspects of theinvention are illustrated further by the following discussion.

[0178] Polynucleotide Assays

[0179] As discussed in some detail supra, this invention is also relatedto the use of LSG polynucleotides to detect complementarypolynucleotides such as, for example, as a diagnostic reagent. Detectionof a mutated form of LSG associated with a dysfunction will provide adiagnostic tool that can add to or define a diagnosis of a disease orsusceptibility to a disease which results from under-expression,over-expression or altered expression of a LSG, such as, for example, asusceptibility to inherited lung cancer.

[0180] Individuals carrying mutations in a human LSG gene may bedetected at the DNA level by a variety of techniques. Nucleic acids fordiagnosis may be obtained from a patient's cells, such as from blood,urine, saliva, tissue biopsy and autopsy material. The genomic DNA maybe used directly for detection or may be amplified enzymatically usingPCR prior to analysis(Saiki et al., Nature, 324: 163-166 (1986)). RNA orcDNA may also be used in a similar manner. As an example, PCR primerscomplementary to a LSG polynucleotide of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7,8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 37 or 38 canbe used to identify and analyze LSG expression and mutations. Forexample, deletions and insertions can be detected by a change in size ofthe amplified product in comparison to the normal genotype. Pointmutations can be identified by hybridizing amplified DNA to radiolabeledLSG RNA or alternatively, radiolabeled LSG antisense DNA sequences.Perfectly matched sequences can be distinguished from mismatchedduplexes by RNase A digestion or by differences in melting temperatures.

[0181] Sequence differences between a reference gene and genes havingmutations also may be revealed by direct DNA sequencing. In addition,cloned DNA segments may be employed as probes to detect specific DNAsegments. The sensitivity of such methods can be greatly enhanced byappropriate use of PCR or another amplification method. For example, asequencing primer is used with double-stranded PCR product or asingle-stranded template molecule generated by a modified PCR. Thesequence determination is performed by conventional procedures withradiolabeled nucleotide or by automatic sequencing procedures withfluorescent-tags.

[0182] Genetic testing based on DNA sequence differences may be achievedby detection of alterations in electrophoretic mobility of DNA fragmentsin gels, with or without denaturing agents. Small sequence deletions andinsertions can be visualized by high resolution gel electrophoresis. DNAfragments of different sequences may be distinguished on denaturingformamide gradient gels in which the mobilities of different DNAfragments are retarded in the gel at different positions according totheir specific melting or partial melting temperatures (see, e.g., Myerset al., Science, 230: 1242 (1985)).

[0183] Sequence changes at specific locations also may be revealed bynuclease protection assays, such as RNase and S1 protection or thechemical cleavage method (e.g., Cotton et al., Proc. Natl. Acad. Sci.,USA, 85: 4397-4401 (1985)).

[0184] Thus, the detection of a specific DNA sequence may be achieved bymethods such as hybridization, RNase protection, chemical cleavage,direct DNA sequencing or the use of restriction enzymes, (e.g.,restriction fragment length polymorphisms (“RFLP”) and Southern blottingof genomic DNA. In addition to more conventional gel-electrophoresis andDNA sequencing, mutations also can be detected by in situ analysis.

[0185] Chromosome Assays

[0186] The LSG sequences of the present invention are also valuable forchromosome identification. There is a need for identifying particularsites on the chromosome and few chromosome marking reagents based onactual sequence data (repeat polymorphisms) are presently available formarking chromosomal location. Each LSG sequence of the present inventionis specifically targeted to and can hybridize with a particular locationon an individual human chromosome. Thus, the LSGs can be used in themapping of DNAs to chromosomes, an important first step in correlatingsequences with genes associated with disease.

[0187] In certain preferred embodiments in this regard, the cDNA hereindisclosed is used to clone genomic DNA of a LSG of the presentinvention. This can be accomplished using a variety of well knowntechniques and libraries, which generally are available commercially.The genomic DNA is used for in situ chromosome mapping using well knowntechniques for this purpose.

[0188] In some cases, sequences can be mapped to chromosomes bypreparing PCR primers (preferably 15-25 bp) from the cDNA. Computeranalysis of the 3′ untranslated region of the gene is used to rapidlyselect primers that do not span more than one exon in the genomic DNA,thus complicating the amplification process. These primers are then usedfor PCR screening of somatic cell hybrids containing individual humanchromosomes. Only those hybrids containing the human gene correspondingto the primer will yield an amplified fragment.

[0189] PCR mapping of somatic cell hybrids is a rapid procedure forassigning a particular DNA to a particular chromosome. Using the presentinvention with the same oligonucleotide primers, sublocalization can beachieved with panels of fragments from specific chromosomes or pools oflarge genomic clones in an analogous manner. Other mapping strategiesthat can similarly be used to map to its chromosome include in situhybridization, prescreening with labeled flow-sorted chromosomes andpreselection by hybridization to construct chromosome specific-cDNAlibraries.

[0190] Fluorescence in situ hybridization (“FISH”) of a cDNA clone to ametaphase chromosomal spread can be used to provide a precisechromosomal location in one step. This technique can be used with cDNAas short as 50 or 60 bp. This technique is described by Verma et al.(HUMAN CHROMOSOMES: A MANUAL OF BASIC TECHNIQUES, Pergamon Press, NewYork (1988)).

[0191] Once a sequence has been mapped to a precise chromosomallocation, the physical position of the sequence on the chromosome can becorrelated with genetic map data. Such data are found, for example, inV. McKusick, MENDELIAN INHERITANCE IN MAN, available on line throughJohns Hopkins University, Welch Medical Library. The relationshipbetween genes and diseases that have been mapped to the same chromosomalregion are then identified through linkage analysis (coinheritance ofphysically adjacent genes).

[0192] Next, it is necessary to determine the differences in the cDNA orgenomic sequence between affected and unaffected individuals. If amutation is observed in some or all of the affected individuals but notin any normal individuals, then the mutation is likely to be thecausative agent of the disease.

[0193] With current resolution of physical mapping and genetic mappingtechniques, a cDNA precisely localized to a chromosomal regionassociated with the disease could be one of between 50 and 500 potentialcausative genes. (This assumes 1 megabase mapping resolution and onegene per 20 kb).

[0194] Polypeptide Assays

[0195] As described in some detail supra, the present invention alsorelates to diagnostic assays such as quantitative and diagnostic assaysfor detecting levels of LSG polypeptide in cells and tissues, andbiological fluids such as blood and urine, including determination ofnormal and abnormal levels. Thus, for instance, a diagnostic assay inaccordance with the present invention for detecting over-expression orunder-expression of a LSG polypeptide compared to normal control tissuesamples may be used to detect the presence of neoplasia. Assaytechniques that can be used to determine levels of a protein, such as aLSG polypeptide of the present invention, in a sample derived from ahost are well-known to those of skill in the art. Such assay methodsinclude radioimmunoassays, competitive-binding assays, Western Blotanalysis and ELISA assays. Among these ELISAs frequently are preferred.

[0196] For example, antibody-sandwich ELISAs are used to detectpolypeptides in a sample, preferably a biological sample. Wells of amicrotiter plate are coated with specific antibodies, at a finalconcentration of 0.2 to 10 μg/ml. The antibodies are either monoclonalor polyclonal and are produced by methods as described herein. The wellsare blocked so that non-specific binding of the polypeptide to the wellis reduced. The coated wells are then incubated for >2 hours at roomtemperature with a sample containing the LSG polypeptide. Preferably,serial dilutions of the sample should be used to validate results. Theplates are then washed three times with deionized or distilled water toremove unbounded polypeptide. Next, 50 μl of specific antibody-alkalinephosphatase conjugate, at a concentration of 25-400 ng, is added andincubated for 2 hours at room temperature. The plates are again washedthree times with deionized or distilled water to remove unboundedconjugate. 4-methylumbelliferyl phosphate (MUP) or p-nitrophenylphosphate (NPP) substrate solution (75 μl) is then added to each welland the plate is incubated 1 hour at room temperature. The reaction ismeasured by a microtiter plate reader. A standard curve is preparedusing serial dilutions of a control sample, and polypeptideconcentration is plotted on the X-axis (log scale) while fluorescence orabsorbance is plotted on the Y-axis (linear scale). The concentration ofthe LSG polypeptide in the sample is interpolated using the standardcurve.

[0197] Antibodies

[0198] As discussed in some detail supra, LSG polypeptides, theirfragments or other derivatives, or analogs thereof, or cells expressingthem can be used as an immunogen to produce antibodies thereto. Theseantibodies can be polyclonal or monoclonal antibodies. The presentinvention also includes chimeric, single chain, and humanizedantibodies, as well as Fab fragments, or the product of an Fabexpression library. Various procedures known in the art may be used forthe production of such antibodies and fragments.

[0199] A variety of methods for antibody production are set forth inCurrent Protocols, Chapter 2.

[0200] For example, cells expressing a LSG polypeptide of the presentinvention can be administered to an animal to induce the production ofsera containing polyclonal antibodies. In a preferred method, apreparation of the secreted protein is prepared and purified to renderit substantially free of natural contaminants. This preparation is thenintroduced into an animal in order to produce polyclonal antisera ofgreater specific activity. The antibody obtained will bind with the LSGpolypeptide itself. In this manner, even a sequence encoding only afragment of the LSG polypeptide can be used to generate antibodiesbinding the whole native polypeptide. Such antibodies can then be usedto isolate the LSG polypeptide from tissue expressing that LSGpolypeptide.

[0201] Alternatively, monoclonal antibodies can be prepared. Examples oftechniques for production of monoclonal antibodies include, but are notlimited to, the hybridoma technique (Kohler, G. and Milstein, C., Nature256: 495-497 (1975), the trioma technique, the human B-cell hybridomatechnique (Kozbor et al., Immunology Today 4: 72 (1983) and (Cole etal., pg. 77-96 in MONOCLONAL ANTIBODIES AND CANCER THERAPY, Alan R.Liss, Inc. (1985). The EBV-hybridoma technique is useful in productionof human monoclonal antibodies.

[0202] Hybridoma technologies have also been described by Khler et al.(Eur. J. Immunol. 6: 511 (1976)) Khler et al. (Eur. J. Immunol. 6: 292(1976)) and Hammerling et al. (in: Monoclonal Antibodies and T-CellHybridomas, Elsevier, N.Y., pp. 563-681 (1981)). In general, suchprocedures involve immunizing an animal (preferably a mouse) with LSGpolypeptide or, more preferably, with a secreted LSGpolypeptide-expressing cell. Such cells may be cultured in any suitabletissue culture medium; however, it is preferable to culture cells inEarle's modified Eagle's medium supplemented with 10% fetal bovine serum(inactivated at about 56° C.), and supplemented with about 10 g/l ofnonessential amino acids, about 1,000 U/ml of penicillin, and about 100μg/ml of streptomycin. The splenocytes of such mice are extracted andfused with a suitable myeloma cell line. Any suitable myeloma cell linemay be employed in accordance with the present invention; however, it ispreferable to employ the parent myeloma cell line (SP20), available fromthe ATCC. After fusion, the resulting hybridoma cells are selectivelymaintained in HAT medium, and then cloned by limiting dilution asdescribed by Wands et al. (Gastroenterology 80: 225-232 (1981).). Thehybridoma cells obtained through such a selection are then assayed toidentify clones which secrete antibodies capable of binding thepolypeptide.

[0203] Alternatively, additional antibodies capable of binding to thepolypeptide can be produced in a two-step procedure using anti-idiotypicantibodies. Such a method makes use of the fact that antibodies arethemselves antigens, and therefore, it is possible to obtain an antibodywhich binds to a second antibody. In accordance with this method,protein specific antibodies are used to immunize an animal, preferably amouse. The splenocytes of such an animal are then used to producehybridoma cells, and the hybridoma cells are screened to identify cloneswhich produce an antibody whose ability to bind to the protein-specificantibody can be blocked by the polypeptide. Such antibodies compriseanti-idiotypic antibodies to the protein specific antibody and can beused to immunize an animal to induce formation of furtherprotein-specific antibodies.

[0204] Techniques described for the production of single chainantibodies (U.S. Pat. No. 4,946,778) can also be adapted to producesingle chain antibodies to immunogenic polypeptide products of thisinvention. Also, transgenic mice, as well as other nonhuman transgenicanimals, may be used to express humanized antibodies to immunogenicpolypeptide products of this invention.

[0205] It will be appreciated that Fab, F(ab′)2 and other fragments ofthe antibodies of the present invention may also be used according tothe methods disclosed herein. Such fragments are typically produced byproteolytic cleavage, using enzymes such as papain (to produce Fabfragments) or pepsin (to produce F(ab′)2 fragments). Alternatively,secreted protein-binding fragments can be produced through theapplication of recombinant DNA technology or through syntheticchemistry.

[0206] For in vivo use of antibodies in humans, it may be preferable touse “humanized” chimeric monoclonal antibodies. Such antibodies can beproduced using genetic constructs derived from hybridoma cells producingthe monoclonal antibodies described above. Methods for producingchimeric antibodies are known in the art (See, for review, Morrison,Science 229: 1202 (1985); Oi et al., BioTechniques 4: 214 (1986);Cabilly et al., U. S. Pat. No. 4,816,567; Taniguchi et al., EP 171496;Morrison et al., EP 173494; Neuberger et al., WO 8601533; Robinson etal., WO 8702671; Boulianne et al., Nature 312: 643 (1984); Neuberger etal., Nature 314: 268 (1985).)

[0207] The above-described antibodies may be employed to isolate or toidentify clones expressing LSG polypeptides or purify LSG polypeptidesof the present invention by attachment of the antibody to a solidsupport for isolation and/or purification by affinity chromatography. Asdiscussed in more detail supra, antibodies specific against a LSG mayalso be used to image tumors, particularly cancer of the lung, inpatients suffering from cancer. Such antibodies may also be usedtherapeutically to target tumors expressing a LSG.

[0208] Preferred exemplary antigenic epitopes of LSGs of the presentinvention which have been identified are depicted below. Theantigenicity index (AI avg) used is Jameson-Wolf. In some embodiment, itmay be preferred to raise antibodies against these regions of the LSGs.positions AI avg length LSG of SEQ ID NO:39 176-220 1.37 45 399-410 1.1812 301-317 1.13 17 370-391 1.13 22 23-34 1.07 12 149-174 1.00 26 51-671.00 17 LSG of SEQ ID NO:42 453-465 1.25 13 399-409 1.25 11 572-584 1.2013 874-887 1.18 14 226-235 1.15 10 30-51 1.09 22 910-920 1.07 11991-1010 1.06 20 655-668 1.06 14 362-373 1.00 12 LSG of SEQ ID NO:44134-160 1.23 27 415-436 1.17 22 485-515 1.16 31 459-474 1.10 16 200-2101.08 11 535-562 1.04 28 91-115 1.04 25 523-532 1.02 10 8-20 1.01 13 LSGof SEQ ID NO:45 563-586 1.19 24 395-408 1.09 14 130-139 1.04 10 117-1271.02 11 165-189 1.01 25 LSG of SEQ ID NO:46 122-137 1.10 16 LSG of SEQID NO:47 1045-1054 1.12 10 845-880 1.10 36 919-945 1.10 27 1376-14181.10 43 144-164 1.10 21 814-835 1.09 22 706-755 1.06 50 401-416 1.05 16445-491 1.04 47 1061-1085 1.03 25 422-442 1.02 21 LSG of SEQ ID NO:48340-362 1.05 23 155-164 1.01 10 228-240 1.00 13 3-14 1.00 12 LSG of SEQID NO:49 189-204 1.08 16 LSG of SEQ ID NO:50 134-143 1.21 10 23-45 1.0123 LSG of SEQ ID NO:51 53-68 1.14 16 LSG of SEQ ID NO:53 367-392 1.32 26491-504 1.07 14 14-35 1.04 22 275-284 1.03 10 208-219 1.03 12 439-4561.02 18 LSG of SEQ ID NO:54 1671-1681 1.35 11 453-465 1.26 13 1748-17591.23 12 1725-1738 1.19 14 1804-1825 1.15 22 1644-1655 1.13 12 1281-12951.12 15 1532-1545 1.11 14 1351-1369 1.07 19 1040-1062 1.06 23 1334-13471.05 14 145-155 1.05 11 1121-1132 1.05 12 1307-1318 1.02 12 1376-14081.02 33 650-660 1.01 11 802-823 1.00 22 714-735 1.00 22 1885-1898 1.0014 1967-1976 1.00 10 LSG of SEQ ID NO:55 297-311 1.31 15 328-344 1.25 1716-25 1.20 10 96-113 1.12 18 381-393 1.12 13 236-250 1.10 15 354-3641.09 11 441-451 1.07 11 274-291 1.00 18 LSG of SEQ ID NO:56 197-210 1.0314 318-328 1.02 11

[0209] LSG Binding Molecules and Assays

[0210] This invention also provides a method for identification ofmolecules, such as receptor molecules, that bind LSGs. Genes encodingproteins that bind LSGs, such as receptor proteins, can be identified bynumerous methods known to those of skill in the art. Examples include,but are not limited to, ligand panning and FACS sorting. Such methodsare described in many laboratory manuals such as, for instance, Coliganet al., Current Protocols in Immunology 1(2): Chapter 5 (1991).

[0211] Expression cloning may also be employed for this purpose. To thisend, polyadenylated RNA is prepared from a cell responsive to a LSG ofthe present invention. A cDNA library is created from this RNA and thelibrary is divided into pools. The pools are then transfectedindividually into cells that are not responsive to a LSG of the presentinvention. The transfected cells then are exposed to labeled LSG. LSGpolypeptides can be labeled by a variety of well-known techniquesincluding, but not limited to, standard methods of radio-iodination orinclusion of a recognition site for a site-specific protein kinase.Following exposure, the cells are fixed and binding of labeled LSG isdetermined. These procedures conveniently are carried out on glassslides. Pools containing labeled LSG are identified as containing cDNAthat produced LSG-binding cells. Sub-pools are then prepared from thesepositives, transfected into host cells and screened as described above.Using an iterative sub-pooling and re-screening process, one or moresingle clones that encode the putative binding molecule, such as areceptor molecule, can be isolated.

[0212] Alternatively a labeled ligand can be photoaffinity linked to acell extract, such as a membrane or a membrane extract, prepared fromcells that express a molecule that it binds, such as a receptormolecule. Cross-linked material is resolved by polyacrylamide gelelectrophoresis (“PAGE”) and exposed to X-ray film. The labeled complexcontaining the ligand-receptor can be excised, resolved into peptidefragments, and subjected to protein microsequencing. The amino acidsequence obtained from microsequencing can be used to design unique ordegenerate oligonucleotide probes to screen cDNA libraries to identifygenes encoding the putative receptor molecule.

[0213] Polypeptides of the invention also can be used to assess LSGbinding capacity of LSG binding molecules, such as receptor molecules,in cells or in cell-free preparations.

[0214] Agonists and Antagonists—Assays and Molecules

[0215] The invention also provides a method of screening compounds toidentify those which enhance or block the action of a LSG on cells. By“compound”, as used herein, it is meant to be inclusive of small organicmolecules, peptides, polypeptides and antibodies as well as any othercandidate molecules which have the potential to enhance or agonize orblock or antagonize the action of LSG on cells. As used herein, anagonist is a compound which increases the natural biological functionsof a LSG or which functions in a manner similar to a LSG, while anantagonist, as used herein, is a compound which decreases or eliminatessuch functions. Various known methods for screening for agonists and/orantagonists can be adapted for use in identifying LSG agonist orantagonists.

[0216] For example, a cellular compartment, such as a membrane or apreparation thereof, such as a membrane-preparation, may be preparedfrom a cell that expresses a molecule that binds a LSG, such as amolecule of a signaling or regulatory pathway modulated by LSG. Thepreparation is incubated with labeled LSG in the absence or the presenceof a compound which may be a LSG agonist or antagonist. The ability ofthe compound to bind the binding molecule is reflected in decreasedbinding of the labeled ligand. Compounds which bind gratuitously, i.e.,without inducing the effects of a LSG upon binding to the LSG bindingmolecule are most likely to be good antagonists. Compounds that bindwell and elicit effects that are the same as or closely related to LSGare agonists. LSG-like effects of potential agonists and antagonists mayby measured, for instance, by determining activity of a second messengersystem following interaction of the candidate molecule with a cell orappropriate cell preparation, and comparing the effect with that of LSGor molecules that elicit the same effects as LSG. Second messengersystems that may be useful in this regard include, but are not limitedto, AMP guanylate cyclase, ion channel or phosphoinositide hydrolysissecond messenger systems.

[0217] Another example of an assay for LSG antagonists is a competitiveassay that combines LSG and a potential antagonist with membrane-boundLSG receptor molecules or recombinant LSG receptor molecules underappropriate conditions for a competitive inhibition assay. LSG can belabeled, such as by radioactivity, such that the number of LSG moleculesbound to a receptor molecule can be determined accurately to assess theeffectiveness of the potential antagonist.

[0218] Potential antagonists include small organic molecules, peptides,polypeptides and antibodies that bind to a LSG polypeptide of theinvention and thereby inhibit or extinguish its activity. Potentialantagonists also may be small organic molecules, a peptide, apolypeptide such as a closely related protein or antibody that binds thesame sites on a binding molecule, such as a receptor molecule, withoutinducing LSG-induced activities, thereby preventing the action of LSG byexcluding LSG from binding.

[0219] Potential antagonists include small molecules which bind to andoccupy the binding site of the LSG polypeptide thereby preventingbinding to cellular binding molecules, such as receptor molecules, suchthat normal biological activity is prevented. Examples of smallmolecules include but are not limited to small organic molecules,peptides or peptide-like molecules.

[0220] Other potential antagonists include antisense molecules.Antisense technology can be used to control gene expression throughantisense DNA or RNA or through triple-helix formation. Antisensetechniques are discussed, for example, in Okano, J. Neurochem. 56: 560(1991); OLIGODEOXYNUCLEOTIDES AS ANTISENSE INHIBITORS OF GENEEXPRESSION, CRC Press, Boca Raton, Fla. (1988). Triple helix formationis discussed in, for instance Lee et al., Nucleic Acids Research 6: 3073(1979); Cooney et al., Science 241: 456 (1988); and Dervan et al.,Science 251: 1360 (1991). The methods are based on binding of apolynucleotide to a complementary DNA or RNA. For example, the 5′ codingportion of a polynucleotide that encodes a mature LSG polypeptide of thepresent invention may be used to design an antisense RNA oligonucleotideof from about 10 to 40 base pairs in length. A DNA oligonucleotide isdesigned to be complementary to a region of the gene involved intranscription thereby preventing transcription and the production of aLSG polypeptide. The antisense RNA oligonucleotide hybridizes to themRNA in vivo and blocks translation of the mRNA molecule into a LSGpolypeptide.

[0221] The oligonucleotides described above can also be delivered tocells such that the antisense RNA or DNA may be expressed in vivo toinhibit production of a LSG.

[0222] Compositions

[0223] The present invention also relates to compositions comprising aLSG polynucleotide or a LSG polypeptide or an agonist or antagonistthereof.

[0224] For example, a LSG polynucleotide, polypeptide or an agonist orantagonist thereof of the present invention may be employed incombination with a non-sterile or sterile carrier or carriers for usewith cells, tissues or organisms, such as a pharmaceutical carriersuitable for administration to a subject. Such compositions comprise,for instance, a media additive or a therapeutically effective amount ofa polypeptide of the invention and a pharmaceutically acceptable carrieror excipient. Such carriers may include, but are not limited to, saline,buffered saline, dextrose, water, glycerol, ethanol and combinationsthereof. The formulation should suit the mode of administration.

[0225] Compositions of the present invention will be formulated anddosed in a fashion consistent with good medical practice, taking intoaccount the clinical condition of the individual patient (especially theside effects of treatment with the polypeptide or other compound alone),the site of delivery, the method of administration, the scheduling ofadministration, and other factors known to practitioners. The “effectiveamount” for purposes herein is thus determined by such considerations.As a general proposition, the total pharmaceutically effective amount ofsecreted polypeptide administered parenterally per dose will be in therange of about 1, μg/kg/day to 10 mg/kg/day of patient body weight,although, as noted above, this will be subject to therapeuticdiscretion. More preferably, this dose is at least 0.01 mg/kg/day, andmost preferably for humans between about 0.01 and 1 mg/kg/day for thehormone. If given continuously, the polypeptide or other compound istypically administered at a dose rate of about 1 μg/kg/hour to about 50mg/kg/hour, either by 1-4 injections per day or by continuoussubcutaneous infusion, for example, using a mini-pump. An intravenousbag solution may also be employed. The length of treatment needed toobserve changes and the interval following treatment for responses tooccur appears to vary depending on the desired effect.

[0226] Pharmaceutical compositions containing the secreted protein ofthe invention are administered orally, rectally, parenterally,intracistemally, intravaginally, intraperitoneally, topically (as bypowders, ointments, gels, drops or transdermal patch), bucally, or as anoral or nasal spray. “Pharmaceutically acceptable carrier” refers to anon-toxic solid, semisolid or liquid filler, diluent, encapsulatingmaterial or formulation auxiliary of any type. The term “parenteral” asused herein refers to modes of administration which include intravenous,intramuscular, intraperitoneal, intrasternal, subcutaneous andintraarticular injection and infusion.

[0227] The polypeptide or other compound is also suitably administeredby sustained-release systems. Suitable examples of sustained-releasecompositions include semipermeable polymer matrices in the form ofshaped articles, e. g., films, or microcapsules. Sustained-releasematrices include polylactides (U.S. Pat. No. 3,773,919 and EP 58481),copolymers of L-glutamic acid and gamma-ethyl-L-glutamate (Sidman, U. etal., Biopolymers 22: 547-556 (1983)), poly (2-hydroxyethyl methacrylate)(R. Langer et al., J. Biomed. Mater. Res. 15: 167-277 (1981), and R.Langer, Chem. Tech. 12: 98-105 (1982)), ethylene vinyl acetate (R.Langer et al.) and poly-D-(−)-3-hydroxybutyric acid (EP 133,988).Sustained-release compositions also include liposomally entrappedpolypeptides. Liposomes containing the polypeptide or other compound areprepared by well known methods (Epstein et al., Proc. Natl. Acad. Sci.USA 82: 3688-3692 (1985); Hwang et al., Proc. Natl. Acad. Sci. USA 77:4030-4034 (1980); EP 52322; EP 36676; EP 88046; EP 143949; EP 142641;Japanese Pat. Appl. 83-118008; U.S. Pat. No. 4,485,045 and 4,544,545;and EP 102324). Ordinarily, the liposomes are of the small (about200-800 Angstroms) unilamellar type in which the lipid content isgreater than about 30 mol. percent cholesterol, the selected proportionbeing adjusted for the optimal therapy.

[0228] For parenteral administration, in one embodiment, the polypeptideor other compound is formulated generally by mixing it at the desireddegree of purity, in a unit dosage injectable form (solution,suspension, or emulsion), with a pharmaceutically acceptable carrier,i.e., one that is non-toxic to recipients at the dosages andconcentrations employed and is compatible with other ingredients of theformulation.

[0229] For example, the formulation preferably does not includeoxidizing agents and other compounds that are known to be deleterious tothe polypeptide or other compound.

[0230] Generally, the formulations are prepared by contacting thepolypeptide or other compound uniformly and intimately with liquidcarriers or finely divided solid carriers or both. Then, if necessary,the product is shaped into the desired formulation. Preferably thecarrier is a parenteral carrier, more preferably a solution that isisotonic with the blood of the recipient. Examples of such carriervehicles include water, saline, Ringer's solution, and dextrosesolution. Non-aqueous vehicles such as fixed oils and ethyl oleate arealso useful herein, as well as liposomes.

[0231] The carrier suitably contains minor amounts of additives such assubstances that enhance isotonicity and chemical stability. Suchmaterials are non-toxic to recipients at the dosages and concentrationsemployed, and include buffers such as phosphate, citrate, succinate,acetic acid, and other organic acids or their salts; antioxidants suchas ascorbic acid; low molecular weight (less than about ten residues)polypeptides, e. g., polyarginine or tripeptides; proteins, such asserum albumin, gelatin, or immunoglobulins; hydrophilic polymers such aspolyvinylpyrrolidone; amino acids, such as glycine, glutamic acid,aspartic acid, or arginine; monosaccharides, disaccharides, and othercarbohydrates including cellulose or its derivatives, glucose, mannose,or dextrins; chelating agents such as EDTA; sugar alcohols such asmannitol or sorbitol; counterions such as sodium; and/or nonionicsurfactants such as polysorbates, poloxamers, or PEG.

[0232] The polypeptide or other compound is typically formulated in suchvehicles at a concentration of about 0.1 mg/ml to 100 mg/ml, preferably1-10 mg/ml, at a pH of about 3 to 8. It will be understood that the useof certain of the foregoing excipients, carriers, or stabilizers willresult in the formation of polypeptide salts or salts of the othercompounds.

[0233] Any polypeptide to be used for therapeutic administration shouldbe sterile. Sterility is readily accomplished by filtration throughsterile filtration membranes (e. g., 0.2 micron membranes). Therapeuticpolypeptide compositions generally are placed into a container having asterile access port, for example, an intravenous solution bag or vialhaving a stopper pierceable by a hypodermic injection needle.

[0234] Polypeptides ordinarily will be stored in unit or multi-dosecontainers, for example, sealed ampules or vials, as an aqueous solutionor as a lyophilized formulation for reconstitution. As an example of alyophilized formulation, 10-ml vials are filled with 5 ml ofsterile-filtered 1 % (w/v) aqueous polypeptide solution, and theresulting mixture is lyophilized. The infusion solution is prepared byreconstituting the lyophilized polypeptide using bacteriostaticWater-for-Injection.

[0235] Kits

[0236] The invention further relates to pharmaceutical packs and kitscomprising one or more containers filled with one or more of theingredients of the aforementioned compositions of the invention.Associated with such container(s) can be a notice in the form prescribedby a governmental agency regulating the manufacture, use or sale ofpharmaceuticals or biological products, reflecting approval by theagency of the manufacture, use or sale of the product for humanadministration.

[0237] Administration

[0238] LSG polypeptides or polynucleotides or other compounds,preferably agonists or antagonists thereof of the present invention maybe employed alone or in conjunction with other compounds, such astherapeutic compounds.

[0239] The pharmaceutical compositions may be administered in anyeffective, convenient manner including, for instance, administration bytopical, oral, anal, vaginal, intravenous, intraperitoneal,intramuscular, subcutaneous, intranasal or intradermal routes amongothers.

[0240] The pharmaceutical compositions generally are administered in anamount effective for treatment or prophylaxis of a specific indicationor indications. In general, the compositions are administered in anamount of at least about 10 g/kg body weight. However, it will beappreciated that optimum dosage will be determined by standard methodsfor each treatment modality and indication, taking into account theindication, its severity, route of administration, complicatingconditions and the like.

[0241] It will be appreciated that conditions caused by a decrease inthe standard or normal expression level of a LSG polypeptide in anindividual can be treated by administering the LSG polypeptide of thepresent invention, preferably in the secreted form, or an agonistthereof. Thus, the invention also provides a method of treatment of anindividual in need of an increased level of a LSG polypeptide comprisingadministering to such an individual a pharmaceutical compositioncomprising an amount of the LSG polypeptide or an agonist thereof toincrease the activity level of the LSG polypeptide in such anindividual. For example, a patient with decreased levels of a LSGpolypeptide may receive a daily dose 0.1-100 μg/kg of a LSG polypeptideor agonist thereof for six consecutive days. Preferably, if a LSGpolypeptide is administered it is in the secreted form.

[0242] Compositions of the present invention can also be administered totreating increased levels of a LSG polypeptide. For example, antisensetechnology can be used to inhibit production of a LSG polypeptide of thepresent invention. This technology is one example of a method ofdecreasing levels of a polypeptide, preferably a secreted form, due to avariety of etiologies, such as cancer. A patient diagnosed withabnormally increased levels of a polypeptide can be administeredintravenously antisense polynucleotides at 0.5, 1.0, 1.5, 2.0 and 3.0mg/kg day for 21 days. This treatment is preferably repeated after a7-day rest period if the treatment was well tolerated. Compositionscomprising an antagonist of a LSG polypeptide can also be administeredto decrease levels of LSG in a patient.

[0243] Gene therapy

[0244] The LSG polynucleotides, polypeptides, agonists and antagoniststhat are polypeptides may be employed in accordance with the presentinvention by expression of such polypeptides in vivo, in treatmentmodalities often referred to as “gene therapy.” Thus, for example, cellsfrom a patient may be engineered with a polynucleotide, such as a DNA orRNA, encoding a polypeptide ex vivo, and the engineered cells then canbe provided to a patient to be treated with the polypeptide. Forexample, cells may be engineered ex vivo by the use of a retroviralplasmid vector containing RNA encoding a polypeptide of the presentinvention. Such methods are well-known in the art and their use in thepresent invention will be apparent from the teachings herein.

[0245] Similarly, cells may be engineered in vivo for expression of apolypeptide in vivo by procedures known in the art. For example, apolynucleotide of the invention may be engineered for expression in areplication defective retroviral vector, as discussed supra. Theretroviral expression construct then may be isolated and introduced intoa packaging cell transduced with a retroviral plasmid vector containingRNA encoding a polypeptide of the present invention such that thepackaging cell now produces infectious viral particles containing thegene of interest. These producer cells may be administered to a patientfor engineering cells in vivo and expression of the polypeptide in vivo.These and other methods for administering a polypeptide of the presentinvention would be apparent to those skilled in the art upon reading theinstant application.

[0246] Retroviruses from which the retroviral plasmid vectors hereinabove mentioned may be derived include, but are not limited to, MoloneyMurine Leukemia Virus, spleen necrosis virus, retroviruses such as RousSarcoma Virus, Harvey Sarcoma Virus, avian leukosis virus, gibbon apeleukemia virus, human immunodeficiency virus, adenovirus,Myeloproliferative Sarcoma Virus, and mammary tumor virus. In oneembodiment, the retroviral plasmid vector is derived from Moloney MurineLeukemia Virus.

[0247] Such vectors will include one or more promoters for expressingthe polypeptide. The selection of a suitable promoter will be apparentto those skilled in the art from the teachings contained herein.However, examples of suitable promoters which may be employed include,but are not limited to, the retroviral LTR, the SV40 promoter, the humancytomegalovirus (CMV) promoter described in Miller et al., Biotechniques7: 980-990 (1989), and eukaryotic cellular promoters such as thehistone, RNA polymerase III, and beta-actin promoters. Other viralpromoters which may be employed include, but are not limited to,adenovirus promoters, thymidine kinase (TK) promoters, and B19parvovirus promoters. Additional promoters which may be used includerespiratory syncytial virus (RSV) promoter, inducible promoters such asthe MMT promoter, the metallothionein promoter, heat shock promoters,the albumin promoter, the ApoAI promoter, human globin promoters, viralthymidine kinase promoters such as the Herpes Simplex thymidine kinasepromoter, retroviral LTRs, the beta-actin promoter, and human growthhormone promoters. The promoter also may be the native promoter whichcontrols the gene encoding the polypeptide.

[0248] The nucleic acid sequence encoding the polypeptide of the presentinvention will be placed under the control of a suitable promoter.

[0249] In one embodiment, the retroviral plasmid vector is employed totransduce packaging cell lines to form producer cell lines. Examples ofpackaging cells which may be transfected include, but are not limitedto, the PE501, PA317, Y-2, Y-AM, PA12, T19-14X, VT-19-17-H2, YCRE,YCRIP, GP+E-86, GP+envAml2, and DAN cell lines as described in Miller,A., Human Gene Therapy 1: 5-14 (1990). The vector may be transduced intothe packaging cells through any means known in the art. Such meansinclude, but are not limited to, electroporation, the use of liposomes,and CaPO₄ precipitation. Alternatively, the retroviral plasmid vectormay be encapsulated into a liposome, or coupled to a lipid, and thenadministered to a host. The producer cell line will generate infectiousretroviral vector particles which are inclusive of the nucleic acidsequence(s) encoding the polypeptides. Such retroviral vector particlesthen may be employed to transduce eukaryotic cells, either in vitro orin vivo. The transduced eukaryotic cells will express the nucleic acidsequence(s) encoding the polypeptide. Eukaryotic cells which may betransduced include, but are not limited to, embryonic stem cells,embryonic carcinoma cells, as well as hematopoietic stem cells,hepatocytes, fibroblasts, myoblasts, keratinocytes, endothelial cells,and bronchial epithelial cells.

[0250] An exemplary method of gene therapy involves transplantation offibroblasts which are capable of expressing a LSG polypeptide or anagonist or antagonist thereof onto a patient. Generally fibroblasts areobtained from a subject by skin biopsy. The resulting tissue is placedin tissue-culture medium and separated into small pieces. Small chunksof the tissue are placed on a wet surface of a tissue culture flask,approximately ten pieces are placed in each flask. The flask is turnedupside down, closed tight and left at room temperature over night. After24 hours at room temperature, the flask is inverted and the chunks oftissue remain fixed to the bottom of the flask and fresh media (e. g.,Ham's F12 media, with 10% FBS, penicillin and streptomycin) is added.The flasks are then incubated at 37° C. for approximately one week. Atthis time, fresh media is added and subsequently changed every severaldays. After an additional two weeks in culture, a monolayer offibroblasts emerge. The monolayer is trypsinized and scaled into largerflasks. pMV-7 (Kirschmeier, P. T. et al., DNA, 7: 219-25 (1988)),flanked by the long terminal repeats of the Moloney murine sarcomavirus, is digested with EcoRI and HindIII and subsequently treated withcalf intestinal phosphatase. The linear vector is fractionated onagarose gel and purified, using glass beads. The cDNA encoding a LSGpolypeptide of the present invention or an agonist or antagonist thereofcan be amplified using PCR primers which correspond to their 5′ and 3′end sequences respectively. Preferably, the 5′ primer contains an EcoRIsite and the 3′ primer includes a HindIII site. Equal quantities of theMoloney murine sarcoma virus linear backbone and the amplified EcoRI andHindIII fragment are added together in the presence of T4 DNA ligase.The resulting mixture is maintained under conditions appropriate forligation of the two fragments. The ligation mixture is then used totransform bacteria HB 101, which are then plated onto agar containingkanamycin for the purpose of confirming that the vector has the gene ofinterest properly inserted. Amphotropic pA317 or GP+aml2 packaging cellsare grown in tissue culture to confluent density in Dulbecco's ModifiedEagles Medium (DMEM) with 10% calf serum (CS), penicillin andstreptomycin. The MSV vector containing the gene is then added to themedia and the packaging cells transduced with the vector. The packagingcells now produce infectious viral particles containing the gene (thepackaging cells are now referred to as producer cells). Fresh media isadded to the transduced producer cells, and subsequently, the media isharvested from a 10 cm plate of confluent producer cells. The spentmedia, containing the infectious viral particles, is filtered through amillipore filter to remove detached producer cells and this media isthen used to infect fibroblast cells. Media is removed from asub-confluent plate of fibroblasts and quickly replaced with the mediafrom the producer cells. This media is removed and replaced with freshmedia. If the titer of virus is high, then virtually all fibroblastswill be infected and no selection is required. If the titer is very low,then it is necessary to use a retroviral vector that has a selectablemarker, such as neo or his. Once the fibroblasts have been efficientlyinfected, the fibroblasts are analyzed to determine whether protein isproduced. The engineered fibroblasts are then transplanted onto thehost, either alone or after having been grown to confluence on cytodex 3microcarrier beads.

[0251] Alternatively, in vivo gene therapy methods can be used to treatLSG related disorders, diseases and conditions. Gene therapy methodsrelate to the introduction of naked nucleic acid (DNA, RNA, andantisense DNA or RNA) sequences into an animal to increase or decreasethe expression of the polypeptide.

[0252] For example, a LSG polynucleotide of the present invention or anucleic acid sequence encoding an agonist or antagonist thereto may beoperatively linked to a promoter or any other genetic elements necessaryfor the expression of the polypeptide by the target tissue. Such genetherapy and delivery techniques and methods are known in the art, see,for example, WO 90/11092, WO 98/11779; U.S. Pat. Nos. 5,693,622,5,705,151, and 5,580,859; Tabata H. et al. (1997) Cardiovasc. Res. 35(3): 470-479, Chao J et al. (1997) Pharmacol. Res. 35 (6): 517-522,Wolff J. A. (1997) Neuromuscul. Disord. 7 (5): 314-318, Schwartz B. etal. (1996) Gene Ther. 3 (5): 405-411, Tsurumi Y. et al. (1996)Circulation 94 (12): 3281-3290 (incorporated herein by reference). Thepolynucleotide constructs may be delivered by any method that deliversinjectable materials to the cells of an animal, such as, injection intothe interstitial space of tissues (heart, muscle, skin, lung, liver,intestine and the like). The polynucleotide constructs can be deliveredin a pharmaceutically acceptable liquid or aqueous carrier.

[0253] The term “naked” polynucleotide, DNA or RNA, refers to sequencesthat are free from any delivery vehicle that acts to assist, promote, orfacilitate entry into the cell, including viral sequences, viralparticles, liposome formulations, lipofectin or precipitating agents andthe like. However, polynucleotides may also be delivered in liposomeformulations (such as those taught in Felgner P. L. et al. (1995) Ann.NY Acad. Sci. 772: 126-139 and Abdallah B. et al. (1995) Biol. Cell 85(1): 1-7) which can be prepared by methods well known to those skilledin the art.

[0254] The polynucleotide vector constructs used in the gene therapymethod are preferably constructs that will not integrate into the hostgenome nor will they contain sequences that allow for replication. Anystrong promoter known to those skilled in the art can be used fordriving the expression of DNA. Unlike other gene therapies techniques,one major advantage of introducing naked nucleic acid sequences intotarget cells is the transitory nature of the polynucleotide synthesis inthe cells. Studies have shown that non-replicating DNA sequences can beintroduced into cells to provide production of the desired polypeptidefor periods of up to six months.

[0255] The polynucleotide construct can be delivered to the interstitialspace of tissues within the an animal, including of muscle, skin, brain,lung, liver, spleen, bone marrow, thymus, heart, lymph, blood, bone,cartilage, pancreas, kidney, gall bladder, stomach, intestine, testis,ovary, uterus, rectum, nervous system, eye, gland, and connectivetissue. Interstitial space of the tissues comprises the intercellularfluid, mucopolysaccharide matrix among the reticular fibers of organtissues, elastic fibers in the walls of vessels or chambers, collagenfibers of fibrous tissues, or that same matrix within connective tissueensheathing muscle cells or in the lacunae of bone. It is similarly thespace occupied by the plasma of the circulation and the lymph fluid ofthe lymphatic channels. Delivery to the interstitial space of muscletissue is preferred. The polynucleotide construct may be convenientlydelivered by injection into the tissues comprising these cells. They arepreferably delivered to and expressed in persistent, non-dividing cellswhich are differentiated, although delivery and expression may beachieved in non-differentiated or less completely differentiated cells,such as, for example, stem cells of blood or skin fibroblasts. In vivomuscle cells are particularly competent in their ability to take up andexpress polynucleotides.

[0256] For the naked polynucleotide injection, an effective dosageamount of DNA or RNA will be in the range of from about 0.05 μg/kg bodyweight to about 50 mg/kg body weight. Preferably the dosage will be fromabout 0.005 mg/kg to about 20 mg/kg and more preferably from about 0.05mg/kg to about 5 mg/kg. Of course, as the artisan of ordinary skill willappreciate, this dosage will vary according to the tissue site ofinjection. The appropriate and effective dosage of nucleic acid sequencecan readily be determined by those of ordinary skill in the art and maydepend on the condition being treated and the route of administration.The preferred route of administration is by the parenteral route ofinjection into the interstitial space of tissues. However, otherparenteral routes may also be used, such as, inhalation of an aerosolformulation particularly for delivery to lungs or bronchial tissues,throat or mucous membranes of the nose. In addition, nakedpolynucleotide constructs can be delivered to arteries duringangioplasty by the catheter used in the procedure.

[0257] The dose response effects of injected polynucleotide in muscle invivo is determined as follows. Suitable template DNA for production ofmRNA coding for polypeptide of the present invention is prepared inaccordance with a standard recombinant DNA methodology. The templateDNA, which may be either circular or linear, is either used as naked DNAor complexed with liposomes. The quadriceps muscles of mice are theninjected with various amounts of the template DNA.

[0258] Five to six week old female and male Balb/C mice are anesthetizedby intraperitoneal injection with 0.3 ml of 2.5% Avertin. A 1.5 cmincision is made on the anterior thigh, and the quadriceps muscle isdirectly visualized. The template DNA is injected in 0.1 ml of carrierin a 1 cc syringe through a 27 gauge needle over one minute,approximately 0.5 cm from the distal insertion site of the muscle intothe knee and about 0.2 cm deep. A suture is placed over the injectionsite for future localization, and the skin is closed with stainlesssteel clips.

[0259] After an appropriate incubation time (e.g., 7 days) muscleextracts are prepared by excising the entire quadriceps. Every fifth 15μm cross-section of the individual quadriceps muscles is histochemicallystained for protein expression. A time course for protein expression maybe done in a similar fashion except that quadriceps from different miceare harvested at different times. Persistence of DNA in muscle followinginjection may be determined by Southern blot analysis after preparingtotal cellular DNA and HIRT supernatants from injected and control mice.

[0260] The results of the above experimentation in mice can be use toextrapolate proper dosages and other treatment parameters in humans andother animals using naked DNA.

[0261] Nonhuman Transgenic Animals

[0262] The LSG polypeptides of the invention can also be expressed innonhuman transgenic animals. Nonhuman animals of any species, including,but not limited to, mice, rats, rabbits, hamsters, guinea pigs, pigs,micro-pigs, goats, sheep, cows and non-human primates, e. g., baboons,monkeys, and chimpanzees, may be used to generate transgenic animals.Any technique known in the art may be used to introduce the transgene(I. e., polynucleotides of the invention) into animals to produce thefounder lines of transgenic animals. Such techniques include, but arenot limited to, pronuclear microinjection (Paterson et al., Appl.Microbiol. Biotechnol. 40: 691-698 (1994); Carver et al., Biotechnology(NY) 11: 1263-1270 (1993); Wright et al., Biotechnology (NY) 9: 830-834(1991); and Hoppe et al., U.S. Pat. No. 4,873,191); retrovirus mediatedgene transfer into germ lines (Van der Putten et al., Proc. Natl. Acad.Sci., USA 82: 6148-6152 (1985)), blastocysts or embryos; gene targetingin embryonic stem cells (Thompson et al., Cell 56: 313-321 (1989));electroporation of cells or embryos (Lo, 1983, Mol. Cell. Biol. 3:1803-1814 (1983)); introduction of the polynucleotides of the inventionusing a gene gun (see, e.g., Ulmer et al., Science 259: 1745 (1993);introducing nucleic acid constructs into embryonic pluripotent stemcells and transferring the stem cells back into the blastocyst; andsperm mediated gene transfer (Lavitrano et al., Cell 57: 717-723(1989)). For a review of such techniques, see Gordon, “TransgenicAnimals,” Intl. Rev. Cytol. 115: 171-229 (1989), which is incorporatedby reference herein in its entirety.

[0263] Any technique known in the art may be used to produce transgenicclones containing polynucleotides of the invention, for example, nucleartransfer into enucleated oocytes of nuclei from cultured embryonic,fetal, or adult cells induced to quiescence (Campell et al., Nature 380:64-66 (1996); Wilmut et al., Nature 385: 810813 (1997)).

[0264] The present invention provides for transgenic animals that carrythe transgene in all their cells, as well as animals which carry thetransgene in some, but not all their cells, i.e., mosaic or chimericanimals. The transgene may be integrated as a single transgene or asmultiple copies such as in concatamers, e. g., head-to-head tandems orhead-to-tail tandems. The transgene may also be selectively introducedinto and activated in a particular cell type by following, for example,the teaching of Lasko et al. (Lasko et al., Proc. Natl. Acad. Sci. USA89: 6232-6236 (1992)). The regulatory sequences required for such acell-type specific activation will depend upon the particular cell typeof interest, and will be apparent to those of skill in the art. When itis desired that the polynucleotide transgene be integrated into thechromosomal site of the endogenous gene, gene targeting is preferred.Briefly, when such a technique is to be utilized, vectors containingsome nucleotide sequences homologous to the endogenous gene are designedfor the purpose of integrating, via homologous recombination withchromosomal sequences, into and disrupting the function of thenucleotide sequence of the endogenous gene. The transgene may also beselectively introduced into a particular cell type, thus inactivatingthe endogenous gene in only that cell type, by following, for example,the teaching of Gu et al. (Science 265: 103-106 (1994)). The regulatorysequences required for such a cell-type specific inactivation willdepend upon the particular cell type of interest, and will be apparentto those of skill in the art.

[0265] Once transgenic animals have been generated, the expression ofthe recombinant gene may be assayed utilizing standard techniques.Initial screening may be accomplished by Southern blot analysis or PCRtechniques to analyze animal tissues to verify that integration of thetransgene has taken place. The level of mRNA expression of the transgenein the tissues of the transgenic animals may also be assessed usingtechniques which include, but are not limited to, Northern blot analysisof tissue samples obtained from the animal, in situ hybridizationanalysis, and reverse transcriptase-PCR (rt-PCR). Samples of transgenicgene-expressing tissue may also be evaluated immunocytochemically orimmunohistochemically using antibodies specific for the transgeneproduct.

[0266] Once the founder animals are produced, they may be bred, inbred,outbred, or crossbred to produce colonies of the particular animal.Examples of such breeding strategies include, but are not limited to:outbreeding of founder animals with more than one integration site inorder to establish separate lines; inbreeding of separate lines in orderto produce compound transgenics that express the transgene at higherlevels because of the effects of additive expression of each transgene;crossing of heterozygous transgenic animals to produce animalshomozygous for a given integration site in order to both augmentexpression and eliminate the need for screening of animals by DNAanalysis; crossing of separate homozygous lines to produce compoundheterozygous or homozygous lines; and breeding to place the transgene ona distinct background that is appropriate for an experimental model ofinterest.

[0267] Transgenic animals of the invention have uses which include, butare not limited to, animal model systems useful in elaborating thebiological function of LSG polypeptides of the present invention,studying conditions and/or disorders associated with aberrant expressionof LSGs, and in screening for compounds effective in ameliorating suchLSG associated conditions and/or disorders.

[0268] Knock-Out Animals

[0269] Endogenous gene expression can also be reduced by inactivating or“knocking out” the gene and/or its promoter using targeted homologousrecombination (e. g., see Smithies et al., Nature 317: 230-234 (1985);Thomas & Capecchi, Cell 51: 503512 (1987); Thompson et al., Cell 5:313-321 (1989); each of which is incorporated by reference herein in itsentirety). For example, a mutant, non-functional LSG polynucleotide ofthe invention (or a completely unrelated DNA sequence) flanked by DNAhomologous to the endogenous LSG polynucleotide sequence (either thecoding regions or regulatory regions of the gene) can be used, with orwithout a selectable marker and/or a negative selectable marker, totransfect cells that express polypeptides of the invention in vivo. Inanother embodiment, techniques known in the art are used to generateknockouts in cells that contain, but do not express the gene ofinterest. Insertion of the DNA construct, via targeted homologousrecombination, results in inactivation of the targeted gene. Suchapproaches are particularly suited in research and agricultural fieldswhere modifications to embryonic stem cells can be used to generateanimal offspring with an inactive targeted gene (e. g., see Thomas &Capecchi 1987 and Thompson 1989, supra). This approach can also beroutinely adapted for use in humans provided the recombinant DNAconstructs are directly administered or targeted to the required site invivo using appropriate viral vectors that will be apparent to those ofskill in the art.

[0270] In further embodiments of the invention, cells that aregenetically engineered to express the LSG polypeptides of the invention,or alternatively, that are genetically engineered not to express the LSGpolypeptides of the invention (e. g., knockouts) are administered to apatient in vivo. Such cells may be obtained from the patient or a MHCcompatible donor and can include, but are not limited to, fibroblasts,bone marrow cells, blood cells (e. g., lymphocytes), adipocytes, musclecells, and endothelial cells. The cells are genetically engineered invitro using recombinant DNA techniques to introduce the coding sequenceof polypeptides of the invention into the cells, or alternatively, todisrupt the coding sequence and/or endogenous regulatory sequenceassociated with the polypeptides of the invention, e. g., bytransduction (using viral vectors, and preferably vectors that integratethe transgene into the cell genome) or transfection procedures,including, but not limited to, the use of plasmids, cosmids, YACs, nakedDNA, electroporation, liposomes, etc.

[0271] The coding sequence of the LSG polypeptides of the invention canbe placed under the control of a strong constitutive or induciblepromoter or promoter/enhancer to achieve expression, and preferablysecretion, of the LSG polypeptides of the invention. The engineeredcells which express and preferably secrete the LSG polypeptides of theinvention can be introduced into the patient systemically, e.g., in thecirculation, or intraperitoneally. Alternatively, the cells can beincorporated into a matrix and implanted in the body, e.g., geneticallyengineered fibroblasts can be implanted as part of a skin graft orgenetically engineered endothelial cells can be implanted as part of alymphatic or vascular graft (see, for example, U.S. Pat. No. 5,399,349and U.S. Pat. No. 5,460,959 each of which is incorporated by referenceherein in its entirety).

[0272] When the cells to be administered are non-autologous or non-MHCcompatible cells, they can be administered using well known techniqueswhich prevent the development of a host immune response against theintroduced cells. For example, the cells may be introduced in anencapsulated form which, while allowing for an exchange of componentswith the immediate extracellular environment, does not allow theintroduced cells to be recognized by the host immune system.

[0273] Transgenic and “knock-out” animals of the invention have useswhich include, but are not limited to, animal model systems useful inelaborating the biological function of LSG polypeptides of the presentinvention, studying conditions and/or disorders associated with aberrantLSG expression, and in screening for compounds effective in amelioratingsuch LSG associated conditions and/or disorders.

[0274] The following nonlimiting example is provided to furtherillustrate the present invention.

EXAMPLE

[0275] The following Example is carried out using standard techniques,which are well known and routine to those of skill in the art, exceptwhere otherwise described in detail. Routine molecular biologytechniques of the following example can be carried out as described instandard laboratory manuals, such as Sambrook et al., MOLECULAR CLONING:A LABORATORY MANUAL, 2nd Ed.; Cold Spring Harbor Laboratory Press, ColdSpring Harbor, N.Y. (1989).

[0276] Introduction and Background for Microarray Analysis

[0277] cDNA microarrays are prepared by high-speed robotic printing ofthousands of distinct cDNAs in an ordered array on glass microscopeslides. They are used to measure the relative abundance of specificsequences in two complex samples (Schena et al, 1995; Shalon et al,1996).

[0278] In the microarray procedure, mRNA is isolated from tissues ofinterest, either from a tumor or control (normal or normal adjacenttissue). mRNA (200-600 ng) from cancer tissue or control is reversetranscribed to incorporate the fluorescent nucleotides Cy5 (red) or Cy3(green), respectively. The two populations of fluorescently labeled cDNAare mixed together and hybridized simultaneously to a microarray bearingapproximately 10,000 cDNA elements in a 2cm×2cm area on a glass slide(Microarrays hybridization service: Incyte Genomics, Fremont, Calif.,USA). After hybridization, the slides are scanned with a scanning laserconfocal microscope.

[0279] The scanned image is used to generate the intensity and localbackground measurements for each spot on the array (GEMtools software,Incyte Genomics). For each spot, representing one EST, the ratio of thenormalized Cy5/Cy3 intensities generates a quantitation of the gene'sexpression in one tissue relative to the control, in this case, theexpression in cancer tissue versus either normal or normal adjacenttissue. For example, a gene that shows a Cancer-Cy5 intensity of 3000and a Normal-Cy3 intensity of 1000 is expressed 3-fold more in cancertissue. Advanced analysis software is used to sort and decipher patternsof gene expression from the data (Cluster and Treeview programs,Stanford University; Eisen et al, 1998; Alizadeh et al, 2000). However,the reproducibility study from Incyte shows that the level of detectabledifferential expression is calculated to be approximately plus or minus1.74. Consequently, any elements with observed ratios greater than orequal to 1.8 between cancer and normal are deemed differentiallyexpressed.

REFERENCES

[0280] 1. Schena, M., D. Shalon, R. W. Davis, and P. O. Brown. 1995.Quantitative monitoring of gene expression patterns with a complementarycDNA microarray. Science 270: 467-470.

[0281] 2. Shalon, D., S. J. Smith, and P. O. Brown. 1996. A DNAMicroarray System for Analyzing Complex DNA samples Using Two-colorFluorescent Probe Hybridization. Genome Research 6:639-645.

[0282] 3. Eisen, P. T. Spellman, P. O. Brown, and D. Botstein. 1998.“Cluster analysis and display of genome-wide expression patterns”. PNAS95: 14863-14868.

[0283] 4. Alizadeh, A. A., et al, 2000. “Distinct types of diffuse largeB-cell lymphoma identified by gene expression profiling.” Nature, 403:503-511.

[0284] 5. GEM Microarray Reproducibility Study. Technical specificationsfrom Incyte Genomics.

[0285] Ling diaDexus Microarray Candidates

[0286] Following is a list of “diaDexus microarray candidates” sequencesfor lung cancer, also referred to herein as lung specific genes or LSGs:Sequences Gene ID/Clone ID/ddxid 1 1040286/2746236/18867 2198406/2639142/12801 3 441298/1877647/8255 4 244318/3032060/7048 5429368/2890670/4002 6 975386/289582/5018 7 480710/1911471/12153 81040699/1899557/13678 9 1040383/1556335/3273 10 108494/3130429/3126 11331878/2445607/3070 12 233442/1959959/18837 13 255993/1670828/7873 14897843/1823610/16315 15 414885/2655867/21009 16 1100375/690306/x 176133/3993331/x 18 257782/3032060A/7048A 19 347005/1911471A/12153A 20332710/3130429A/3126A 21 255828/2445607A/3070A 22 328565/3993331A/x

[0287] Table 1 depicts numbers which are ratios indicating the levels ofexpression of the Clone IDs in the cancer tissue sample (labeled withCy5) relative to the normal tissue, or the normal adjacent tissuecontrol (labeled with Cy3) used in that experiment. The Cy5/Cy3 ratio ofthe normalized fluorescent intensities in each channel is used as ameasure of relative gene expression. A positive number representsoverexpression in cancer relative to the normal control. A negativenumber represents higher expression in the normal adjacent samplecompared to the cancer tissue sample used in that experiment. X means noexperiment was performed for the particular tissue sample. TABLE 1LN.A143 LN.A160 LN.A182 CloneID Vs. Apool Vs. Apool Vs. Apool 27462365.3 2.6 1.5 2639142 1.9 1.7 x 1877647 1.6 1.5 x 3032060 2.1 1.2 x2890670 2.6 1.0 2.7 289582 1.5 1.1 x LN.A213 LN.A288 LN.A323 CloneID Vs.Apool Vs. Apool Vs. Apool 2746236 4.6 8.9 3.7 2639142 2.9 x 4.5 18776472   x 5.0 3032060 2.3 x 4.8 2890670 x 4.8 4.6 289582 2.4 x 1.8 LN.A339LN.A345 CloneID Vs. Apool Vs. Apool 2746236 2.9 2.9 2639142 1.7 1.21877647 1.9 1.2 3032060 1.2 1.2 2890670 2.4 1.9 289582 2.4 1.9

[0288] Absolute values greater than or equal to 1.8 are considered to beabove background levels, and are, therefore significant (Source: IncyteGenomics: GEM microarray technical specifications). The relative levelsof expression in Table 1 show that Clone ID 2746236 mRNA expression ishigher than background in 7 of the cancer tissue samples out of a totalof 8 experiments. Clone ID 2639142 mRNA expression is higher thanbackground in 3 of the cancer tissue samples out of a total of 6experiments. Clone ID 1877647 mRNA expression is higher than backgroundin 3 of the cancer tissue samples out of a total of 6 experiments. CloneID 3032060 mRNA expression is higher than background in 3 of the cancertissue samples out of a total of 6 experiments. Clone ID 2890670 mRNAexpression is higher than background in 5 of the cancer tissue samplesout of a total of 6 experiments. Clone ID 289582 mRNA expression ishigher than background in 4 of the cancer tissue samples out of a totalof 6 experiments.

[0289] An additional 16 clones have also been identified by the sametype of experiments. These additional clones all show from 30% to 80 %overexpression in cancer tissue samples. The sequences of these LSGs arealso disclosed herein.

[0290] Semi-Quantitative Polymerase Chain Reaction

[0291] Semi-quantitative Polymerase Chain Reaction (SQ-PCR) is a methodthat utilizes end point PCR on serial dilutions of cDNA samples in orderto determine relative expression patterns of genes of interest inmultiple samples. Using random hexamer primed Reverse Transcription (RT)cDNA panels are created from total RNA samples. Gene specific primersare then used to amplify fragments using Polymerase Chain Reaction (PCR)technology from four 10× serial cDNA dilutions in duplicate. Relativeexpression levels of 0, 1, 10, 100 and 1000 are used to evaluate geneexpression. A positive reaction in the most dilute sample indicates thehighest relative expression value. This is determined by analysis of thesample reactions on a 2-4% agarose gel. The tissue samples used include12 normal, 12 cancer and 6 pairs tissue specific cancer and matchingsamples.

[0292] Of the list of “diaDexus microarray candidates” sequences forlung cancer, the following sequences were analyzed by semi-quantitativePCR and found to be upregulated in lung adenocarcinoma/carcinoma. SQlngExample# SEQ ID NO: Gene ID Clone ID ddxid code 1 1 1040286 274623618867 Sqlng042 2 3 441298 1877647 8255 Sqlng040 3 11 331878 2445607 3070Sqlng046 4 22 328565 3993331A x Sqlng050

Example 1 SEQ ID NO:1

[0293] Semi quantitative PCR was done using the following primers:Sqlng042 forward: 5′ CCAGAGCCCAAATCTTGTGAC 3′ (SEQ ID NO:23) Sqclng042reverse: 5′ GCGGCTTTGTCTTGGCATTA 3′ (SEQ ID NO:24)

[0294] Table 2 shows absolute numbers which are relative levels ofexpression of Sqlng042 in 12 normal samples from 12 different tissues.These RNA samples are individual samples or are commercially availablepools, originated by pooling samples of a particular tissue fromdifferent individuals. Using Polymerase Chain Reaction (PCR) technologyexpression levels were analyzed from four 10× serial cDNA dilutions induplicate. Relative expression levels of 0, 1, 10, 100 and 1000 are usedto evaluate gene expression. A positive reaction in the most dilutesample indicates the highest relative expression value. TABLE 2 TissueNormal Breast 1000 Colon 1000 Endometruim 1000 Kidney 1000 Liver 10 Lung1000 Ovary 1000 Prostate 100 Small Intestine 1000 Stomach 1000 Testis1000 Uterus 100

[0295] Relative levels of expression in Table 2 show that normal breast,colon, endometrium, kidney, lung, ovary, small intestine, stomach andtestis show high expression of Sqlng042. Moderate levels of expressionare apparent in prostate and uterus. Low levels of expression areapparent in normal liver.

[0296] Table 3 shows absolute numbers which are relative levels ofexpression of Sqlng042 in 12 cancer samples from 12 different tissues.Using Polymerase Chain Reaction (PCR) technology expression levels wereanalyzed from four 10× serial cDNA dilutions in duplicate. Relativeexpression levels of 0, 1, 10, 100 and 1000 are used to evaluate geneexpression. A positive reaction in the most dilute sample indicates thehighest relative expression value. TABLE 3 Tissue Cancer bladder 1000breast 1000 colon 1000 kidney 1 liver 100 lung 1000 ovary 1 pancreas1000 prostate 10 stomach 1000 testes 1 uterus 1000

[0297] Relative levels of expression in Table 3 show that Sqlng042 isexpressed in low levels in kidney, ovary, and testis carcinomas.Sqlng042 is expressed in high levels in other tissue carcinomas.

[0298] Table 4 shows absolute numbers which are relative levels ofexpression of Sqlng042 in 6 lung cancer matching samples. A matchingpair is formed by mRNA from the cancer sample for a particular tissueand mRNA from the normal adjacent sample for that same tissue from thesame individual.

[0299] Using Polymerase Chain Reaction (PCR) technology expressionlevels were analyzed from four 10× serial cDNA dilutions in duplicate.Relative expression levels of 0, 1, 10, 100 and 1000 are used toevaluate gene expression. A positive reaction in the most dilute sampleindicates the highest relative expression value TABLE 4 Sample ID TissueCancer NAT 9702C115RB lung 1 1 9502C032 lung 1000 1000 8894A lung 1 10009704C060RA lung 1 1 11145B lung 1 1000 9502C109R lung 1000 1000

[0300] Relative levels of expression in Table 4 show that Sqlng042 isexpressed in high levels in two of the six lung cancer samples. However,high levels of expression was observed in the matching normal adjacenttissue (NAT).

Example 2 SEQ ID NO:3

[0301] Semi quantitative PCR was done using the following primers:Sqlng040 forward: 5′ ATTGCCATCCCAGTGACAGTG 3′ (SEQ ID NO:25) Sqclng040reverse: 5′ TTGGGAGATGTGGGTGATGAG 3′ (SEQ ID NO:26)

[0302] Table 5 shows absolute numbers which are relative levels ofexpression of Sqlng040 in 12 normal samples from 12 different tissues.These RNA samples are individual samples or are commercially availablepools, originated by pooling samples of a particular tissue fromdifferent individuals. Using Polymerase Chain Reaction (PCR) technologyexpression levels were analyzed from four 10× serial cDNA dilutions induplicate. Relative expression levels of 0, 1, 10, 100 and 1000 are usedto evaluate gene expression. A positive reaction in the most dilutesample indicates the highest relative expression value. TABLE 5 TissueNormal Breast 0 Colon 0 Endometrium 1 Kidney 0 Liver 0 Lung 10 Ovary 1Prostate 10 Small Intestine 1 Stomach 1 Testis 100 Uterus 1

[0303] Relative levels of expression in Table 5 show that normal lungand prostate show moderate expression of Sqlng040o. High levelexpression is only apparent in testis. Low levels of expression areapparent in endometrium, ovary, small intestine and uterus.

[0304] Table 6 shows absolute numbers which are relative levels ofexpression of Sqlng040 in 12 cancer samples from 12 different tissues.Using Polymerase Chain Reaction (PCR) technology expression levels wereanalyzed from four 10× serial cDNA dilutions in duplicate. Relativeexpression levels of 0, 1, 10, 100 and 1000 are used to evaluate geneexpression. A positive reaction in the most dilute sample indicates thehighest relative expression value. TABLE 6 Tissue Cancer bladder 0breast 10 colon 0 kidney 10 liver 0 lung 100 ovary 100 pancreas 100prostate 10 stomach 10 testes 10 uterus 10

[0305] Relative levels of expression in Table 6 show that Sqlng040 isexpressed in moderate to high levels in breast, kidney, lung, ovary,pancreas, prostate, stomach, testis and uterus carcinomas.

[0306] Table 7 shows absolute numbers which are relative levels ofexpression of Sqlng040 in 6 lung cancer matching samples. A matchingpair is formed by mRNA from the cancer sample for a particular tissueand mRNA from the normal adjacent sample for that same tissue from thesame individual. Using Polymerase Chain Reaction (PCR) technologyexpression levels were analyzed from four 10× serial cDNA dilutions induplicate. Relative expression levels of 0, 1, 10, 100 and 1000 are usedto evaluate gene expression. A positive reaction in the most dilutesample indicates the highest relative expression value. TABLE 7 SampleID Tissue Cancer NAT 9702C115RB lung 100 10 9502C032 lung 100 1 8894Alung 10 0 9704C060RA lung 10 10 11145B lung 10 100 9502C109R lung 100 10

[0307] Relative levels of expression in Table 7 show that Sqlng040 isexpressed in moderate levels in four of the six lung cancer samplescompared with the expression in the matching normal adjacent tissue(NAT).

Example 3 SEQ ID NO:11

[0308] Semi quantitative PCR was done using the following primers:Sqlng046 forward: 5′ CCTGCCCTGGTATGTTTTTCTT 3′ (SEQ ID NO:27) Sqlng046reverse: 5′ CAGCCCACAAATGCCTTCTAC 3′ (SEQ ID NO:28)

[0309] Table 8 shows absolute numbers which are relative levels ofexpression of Sqlng046 in 12 normal samples from 12 different tissues.These RNA samples are individual samples or are commercially availablepools, originated by pooling samples of a particular tissue fromdifferent individuals. Using Polymerase Chain Reaction (PCR) technologyexpression levels were analyzed from four 10× serial cDNA dilutions induplicate. Relative expression levels of 0, 1, 10, 100 and 1000 are usedto evaluate gene expression. A positive reaction in the most dilutesample indicates the highest relative expression value. TABLE 8 TissueNormal Breast 0 Colon 10 Endometrium 1 Kidney 10 Liver 1 Lung 10 Ovary10 Prostate 0 Small Intestine 0 Stomach 0 Testis 10 Uterus 1

[0310] Relative levels of expression in Table 8 show that normal colon,kidney, lung, and ovary show moderate expression of Sqlng046. Low levelsof expression are apparent in endometrium and liver. No expression isapparent in other tissues.

[0311] Table 9 shows absolute numbers which are relative levels ofexpression of Sqlng046 in 12 cancer samples from 12 different tissues.Using Polymerase Chain Reaction (PCR) technology expression levels wereanalyzed from four 10× serial cDNA dilutions in duplicate. Relativeexpression levels of 0, 1, 10, 100 and 1000 are used to evaluate geneexpression. A positive reaction in the most dilute sample indicates thehighest relative expression value. Table 9: Tissue Cancer bladder 1breast 1 colon 0 kidney 1 liver 1 lung 0 ovary 0 pancreas 10 prostate 0stomach 1 testes 1 uterus 1

[0312] Relative levels of expression in Table 9 show that Sqlng046 isexpressed in low levels in bladder, breast, kidney, liver, stomach,testis and uterus carcinomas. Sqlng046 is expressed in moderate levelsonly in pancreatic carcinoma.

[0313] Table 10 shows absolute numbers which are relative levels ofexpression of Sqlng046 in 6 lung cancer matching samples. A matchingpair is formed by mRNA from the cancer sample for a particular tissueand mRNA from the normal adjacent sample for that same tissue from thesame individual. Using Polymerase Chain Reaction (PCR) technologyexpression levels were analyzed from four 10× serial cDNA dilutions induplicate. Relative expression levels of 0, 1, 10, 100 and 1000 are usedto evaluate gene expression. A positive reaction in the most dilutesample indicates the highest relative expression value.

[0314] Table 10: Sample ID Tissue Cancer NAT 9702C115RB lung 10 109502C032 lung 100 100 8894A lung 10 1 9704C060RA lung 10 10 11145B lung1 10 9502C109R lung 100 1

[0315] Relative levels of expression in Table 10 show that Sqlng046 isexpressed in higher levels in two of the six lung cancer samplescompared with the expression in matching normal adjacent tissue (NAT).

Example 4 SEQ ID NO:22

[0316] Semi quantitative PCR was done using the following primers:Sqlng050 forward: 5′ CCACTAGGATTATTTCCAGCATAA 3′ (SEQ ID NO:29)Sqclng050 reverse: 5′ GGTGTGAAAATATCTGGTCCACTT 3′ (SEQ ID NO:30)

[0317] Table 12 shows absolute numbers which are relative levels ofexpression of Sqlng050 in 12 normal samples from 12 different tissues.These RNA samples are individual samples or are commercially availablepools, originated by pooling samples of a particular tissue fromdifferent individuals. Using Polymerase Chain Reaction (PCR) technologyexpression levels were analyzed from four 10× serial cDNA dilutions induplicate. Relative expression levels of 0, 1, 10, 100 and 1000 are usedto evaluate gene expression. A positive reaction in the most dilutesample indicates the highest relative expression value. Table 12: TissueNormal Breast 100 Colon 1000 Endometrium 100 Kidney 100 Liver 100 Lung100 Ovary 1000 Prostate 1000 Small Intestine 100 Stomach 100 Testis 10Uterus 100

[0318] Relative levels of expression in Table 12 show that normal colon,ovary, and prostate show high expression of Sqlng050. Moderate levels ofexpression are apparent in breast, endometrium, kidney, liver, lung,small intestine, stomach and uterus. Low levels of expression areapparent in normal testis.

[0319] Table 13 shows absolute numbers which are relative levels ofexpression of Sqlng050 in 12 cancer samples from 12 different tissues.Using Polymerase Chain Reaction (PCR) technology expression levels wereanalyzed from four 10× serial cDNA dilutions in duplicate. Relativeexpression levels of 0, 1, 10, 100 and 1000 are used to evaluate geneexpression. A positive reaction in the most dilute sample indicates thehighest relative expression value. Tissue Cancer bladder 10 breast 100colon 100 kidney 10 liver 100 lung 100 ovary 100 pancreas 100 prostate100 stomach 100 testes 100 uterus 1000

[0320] Relative levels of expression in Table 13 show that Sqlng050 isexpressed in low to moderate levels in 11 out of 12 different tissuecarcinomas. Sqlng050 is only expressed in high level in uteruscarcinoma.

[0321] Table 14 shows absolute numbers which are relative levels ofexpression of Sqlng050 in 6 lung cancer matching samples. A matchingpair is formed by mRNA from the cancer sample for a particular tissueand mRNA from the normal adjacent sample for that same tissue from thesame individual. Using Polymerase Chain Reaction (PCR) technologyexpression levels were analyzed from four 10× serial cDNA dilutions induplicate. Relative expression levels of 0, 1, 10, 100 and 1000 are usedto evaluate gene expression. A positive reaction in the most dilutesample indicates the highest relative expression value. Table 14: SampleID Tissue Cancer NAT 9702C115RB lung 100 100 9502C032 lung 1000 10008894A lung 100 1 9704C060RA lung 100 10 11145B lung 100 1000 9502C109Rlung 100 10

[0322] Relative levels of expression in Table 14 show that Sqlng050 isexpressed in higher levels in three of the six lung cancer samplescompared to the expression level in the matching normal adjacent tissue(NAT).

[0323] Relative Quantitation of Gene Expression

[0324] Real-Time quantitative PCR with fluorescent Taqman probes is aquantitation detection system utilizing the 5′-3′ nuclease activity ofTaq DNA polymerase. The method uses an internal fluorescentoligonucleotide probe (Taqman) labeled with a 5′ reporter dye and adownstream, 3′ quencher dye. During PCR, the 5′-3′ nuclease activity ofTaq DNA polymerase releases the reporter, whose fluorescence can then bedetected by the laser detector of the Model 7700 Sequence DetectionSystem (PE Applied Biosystems, Foster City, Calif., USA).

[0325] Amplification of an endogenous control is used to standardize theamount of sample RNA added to the reaction and normalize for ReverseTranscriptase (RT) efficiency.

[0326] Either cyclophilin, glyceraldehyde-3-phosphate dehydrogenase(GAPDH) or 18S ribosomal RNA (rRNA) is used as this endogenous control.To calculate relative quantitation between all the samples studied, thetarget RNA levels for one sample were used as the basis for comparativeresults (calibrator). Quantitation relative to the “calibrator” can beobtained using the standard curve method or the comparative method (UserBulletin #2: ABI PRISM 7700 Sequence Detection System).

[0327] The tissue distribution and the level of the target gene wereexamined for every example in normal and cancer tissue. Total RNA wasextracted from normal tissues, cancer tissues, and from cancers and thecorresponding matched adjacent tissues. Subsequently, first strand cDNAwas prepared with reverse transcriptase and the polymerase chainreaction was done using primers and Taqman probe specific to each targetgene. The results are analyzed using the ABI PRISM 7700 SequenceDetector. The absolute numbers are relative levels of expression of thetarget gene in a particular tissue compared to the calibrator tissue.

Example 1 SEQ ID NO: 3

[0328] Table 15 shows absolute numbers which are relative levels ofexpression of the LSG of SEQ ID NO:3 in 24 normal different tissues. Allthe values are compared to normal small intestine (calibrator). TheseRNA samples are commercially available pools, originated by poolingsamples of a particular tissue from different individuals. Table 15:Tissue NORMAL Adrenal Gland 0.56 Bladder 0.03 Brain 2.57 Cervix 0.42Colon 0.33 Endometrium 5.12 Esophagus 0.06 Heart 0.08 Kidney 1.2 Liver1.38 Lung 5.54 Mammary Gland 3.96 Muscle 0.44 Ovary 1.29 Pancreas 7.94Prostate 5.21 Rectum 1.36 Small Intestine 1 Spleen 36.89 Stomach 2.8Testis 10.16 Thymus 179.15 Trachea 3.08 Uterus 1.04

[0329] The relative levels of expression in Table 15 show that mRNAexpression of the LSG of SEQ ID NO:3 is very high in thymus (179.15)compared with all the other normal tissues analyzed. The expressionlevel of the LSG of SEQ ID NO:3 is moderate in normal lung. Smallintestine, the calibrator, has a relative expression level of 1. Theseresults demonstrated that mRNA expression of the LSG of SEQ ID NO:3 isrelatively specific for lung.

[0330] The absolute numbers in Table 15 were obtained analyzing pools ofsamples of a particular tissue from different individuals. They can notbe compared to the absolute numbers originated from RNA obtained fromtissue samples of a single individual in Table 16.

[0331] Table 16 shows absolute numbers which are relative levels ofexpression of the LSG of SEQ ID NO:3 in 79 pairs of matching samples and2 normal blood samples. All the values are compared to normal smallintestine (calibrator). A matching pair is formed by mRNA from thecancer sample for a particular tissue and mRNA from the normal adjacentsample for that same tissue from the same individual.

[0332] Table 16: MATCHING Sample Cancer NORMAL ID Type Tissue NORMALCANCER ADJACENT Lng60L Adenocarcinoma Lung 1 1.32 0.95 Lng143LAdenocarcinoma Lung 2 9.29 0.96 Lng60XL Adenocarcinoma Lung 3 41.5 13.18LngAC82 Adenocarcinoma Lung 4 60.97 2.04 LngAC88 Adenocarcinoma Lung 550.21 31.89 LngAC66 Adenocarcinoma Lung 6 1.42 0.72 LngAC69Adenocarcinoma Lung 7 2.3 0.73 LngAC11 Adenocarcinoma Lung 8 2.41 1.95LngAC32 Adenocarcinoma Lung 9 3.9 0.69 LngAC94 Adenocarcinoma Lung 102.65 0.77 LngAC90 Adenocarcinoma Lung 11 16.85 0.57 Lng223LAdenocarcinoma Lung 12 1.48 0.06 LngAC39 Adenocarcinoma Lung 13 139.11.52 LngBR26 Bronchio-alveolar Lung 14 41.79 8.57 carcinoma LngBA641Bronchio-alveolar Lung 15 37.14 16 carcinoma LngSQ45 Squamous cell Lung16 4.92 4.01 carcinoma LngSQ14 Squamous cell Lung 17 7.06 15.19carcinoma LngSQ9X Squamous cell Lung 18 38.32 1.78 carcinoma LngSQ56Squamous cell Lung 19 55.72 33.01 carcinoma LngSQ80 Squamous cell Lung20 34.42 4.3 carcinoma LngSQ32 Squamous cell Lung 21 69.55 21.86carcinoma LngSQ16 Squamous cell Lung 22 1.7 0.22 carcinoma LngSQ79Squamous cell Lung 23 4.71 3.04 carcinoma Lng47XQ Squamous cell Lung 2435.26 1.42 carcinoma LngBR94 Squamous cell Lung 25 138.62 0.19 carcinomaLngC20X Squamous cell Lung 26 3.05 0.18 carcinoma LngSQ44 Squamous cellLung 27 7.06 3.97 carcinoma Lng90X Squamous cell Lung 28 1.49 0.66carcinoma LngSQ43 Squamous cell Lung 29 97.01 1.71 carcinoma LngLC71Large cell Lung 30 27.86 16.22 carcinoma LngLC109 Large cell Lung 31102.89 20.25 carcinoma LngLC80 Large cell Lung 32 34.66 10.13 carcinomaLng77L Large cell Lung 33 1.03 9.22 carcinoma Lng315L Lung carcinomaLung 34 36.25 50.39 Lng528L Lung carcinoma Lung 35 21.48 6.54 Lng75XCMetastatic from Lung 36 3.53 4.55 Osteogenic Sarcoma LngMT67 Metastaticfrom Lung 37 8.2 3.97 renal cell cancer LngMT71 Metastatic from Lung 3813.93 19.23 melanoma Bld46XK Bladder 1 0 0 BldTR14 Bladder 2 1.57 0.78B5 Blood 1 154.34 B6 Blood 2 177.91 CvxKS52 Cervix 1 11.96 2.27 CvxKS83Cervix 2 92.09 8.66 ClnAS43 Colon 1 4.03 0.29 ClnAS45 Colon 2 0.28 0.17ClnAS46 Colon 3 0.38 0.59 Cln AS67 Colon 4 0.62 1.78 Cln AS89 Colon 50.09 0.05 Endo28XA Endometrium 1 15.51 4.77 Endo10479 Endometrium 2 247.14 Endo68X Endometrium 3 13.13 14.42 Kid10XD Kidney 1 3.07 2.07Kid109XD Kidney 2 8.22 7.24 Liv15XA Liver 1 0.17 0.09 Liv174L Liver 20.15 0.32 Mam355 Mammary 1 2.63 0.15 Mam173M Mammary 2 6.87 7.67 Mam220Mammary 3 0.29 0.87 Mam976M Mammary 4 0.19 0.91 Ovr180B Ovary 1 25.72 0OvrA084 Ovary 2 2.7 1.97 Pan77X Pancreas 1 8.11 3.25 Pan92X Pancreas 227.28 21.78 Pro101XB Prostate 1 6.99 4.68 Pro109XB Prostate 2 1.42 1.16Pro125XB Prostate 3 2.24 1.71 Pro13XB Prostate 4 0.41 1.59 Skn39A Skin 13.71 0.35 Skn816S Skin 2 25.81 0.34 SmInt21XA Sm. Int. 1 4.35 1.17SmIntH89 Sm. Int. 2 13.93 3.16 Sto115S Stomach 1 4.59 5.17 Sto2645Stomach 2 6.39 4.16 Sto288S Stomach 3 5.01 0.46 Thr270T Thyroid 1 6.394.58 Thr939T Thyroid 2 0.86 1.55 Tst647T Testis 1 2.49 0.43 Tst663TTestis 2 9.16 3.89 Utr135XO Uterus 1 0.34 0.43 Utr141XO Uterus 2 2.510.63

[0333] In the analysis of matching samples, the higher levels ofexpression were in lung showing a high degree of tissue specificity forlung tissue. These results confirm the tissue specificity resultsobtained with normal pooled samples (Table 15).

[0334] Furthermore, the levels of mRNA expression in cancer samples andthe isogenic normal adjacent tissue from the same individual werecompared. This comparison provides an indication of specificity for thecancer stage (e.g. higher levels of mRNA expression in the cancer samplecompared to the normal adjacent). Table 16 shows overexpression of theLSG of SEQ ID NO:3 in 26 lung cancer tissues compared with theirrespective normal adjacent (lung samples #2, 3, 4, 5, 6, 7, 9, 10, 11,12, 14, 15, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 29, 30, 31, and 32).There is overexpression in the cancer tissue for 68% of the lungmatching samples tested (total of 38 lung matching samples).

[0335] Altogether, the relative high level of lung tissue specificity,plus the mRNA overexpression in 68% of the lung carcinoma matchingsamples tested are believed to make the LSG of SEQ ID NO:3 a gooddiagnostic marker for lung cancer.

[0336] Primers used for expression analysis are: Forward5′ AGCCATTGCCATCCCAGT 3′ (SEQ ID NO:31) Reverse 5′ ATGTTCTTCACGCTCTTCGC3′ (SEQ ID NO:32) Probe 5′ AGGAAGTGCTGGAAGAGGCTGGCT 3′ (SEQ ID NO:33)

Example 2 SEQ ID NO: 15

[0337] Table 17 shows absolute numbers which are relative levels ofexpression of the LSG of SEQ ID NO:15 in 24 normal different tissues.All the values are compared to normal brain (calibrator). These RNAsamples are commercially available pools, originated by pooling samplesof a particular tissue from different individuals.

[0338] Table 17: Tissue NORMAL Adrenal Gland 67.65 Bladder 39.67 Brain1.00 Cervix 677.93 Colon 1287.18 Endometrium 162.58 Esophagus 1034.70Heart 4.81 Kidney 25.02 Liver 194.01 Lung 4705.07 Mammary Gland 840.44Muscle 12.91 Ovary 608.87 Pancreas 20.89 Prostate 858.10 Rectum 4435.87Small Intestine 2149.82 Spleen 5595.30 Stomach 14115.57 Testis 64.67Thymus 2187.40 Trachea 2866.35 Uterus 193.34

[0339] The relative levels of expression in Table 17 show that mRNAexpression of the LSG of SEQ ID NO:15 is very high in stomach (14115.57)compared with all the other normal tissues analyzed. Expression levelsof this LSG are moderate in normal lung (4705.07) Brain, the calibrator,has a relative expression level of 1. These results demonstrate thatmRNA expression of the LSG of SEQ ID NO:15 is relatively specific forlung.

[0340] The absolute numbers in Table 17 were obtained analyzing pools ofsamples of a particular tissue from different individuals. They can notbe compared to the absolute numbers originated from RNA obtained fromtissue samples of a single individual in Table 18. Table 18: MATCHINGSample Cancer NORMAL ID Type Tissue NORMAL CANCER ADJACENT Lng60LAdenocarcinoma Lung 1 18561.17 5732.70 Lng143L Adenocarcinoma Lung 228.54 1.57 LngAC66 Adenocarcinoma Lung 3 16555.24 3408.69 LngAC69Adenocarcinoma Lung 4 18116.29 1891.09 LngAC11 Adenocarcinoma Lung 54389.98 5732.70 LngAC32 Adenocarcinoma Lung 6 18179.19 10015.87 LngAC94Adenocarcinoma Lung 7 10623.71 309.76 Lng223L Adenocarcinoma Lung 88393.17 491.14 LngBR26 Bronchio-alveolar Lung 9 13.98 20.68 carcinomaLngBA641 Bronchio-alveolar Lung 10 34.78 10.13 carcinoma LngSQ45Squamous cell Lung 11 9184.59 8995.58 carcinoma LngSQ14 Squamous cellLung 12 2.82 32.11 carcinoma LngSQ80 Squamous cell Lung 13 68.12 4.07carcinoma LngSQ16 Squamous cell Lung 14 3373.43 86.22 carcinoma LngSQ79Squamous cell Lung 15 19215.37 81245.48 carcinoma Lng9OX Squamous cellLung 16 5.19 1.14 carcinoma LngSQ43 Squamous cell Lung 17 24.17 2.12carcinoma LngLC71 Large cell Lung 18 67.42 25.37 carcinoma LngLC109Large cell Lung 19 12.38 3.96 carcinoma LngMT71 Metastatic from Lung 2013.00 9.45 melanoma Bld46XK Bladder 1 131.60 5.90 BldTR14 Bladder 28306.36 7009.03 CvxKS52 Cervix 1 24.85 8.91 ClnAS43 Colon 1 1590.218335.19 ClnAS45 Colon 2 1458.23 1820.35 ClnAS46 Colon 3 2418.67 3019.30ClnAS67 Colon 4 365.82 823.14 ClnAS89 Colon 5 2304.12 75.32 Endo28XAEndometrium 1 10.70 0.49 Kid10XD Kidney 1 0.38 0.21 Liv15XA Liver 119.16 115.76 Mam355 Mammary 1 16.56 0.18 Pan77X Pancreas 1 0.15 0.07Pro101XB Prostate 1 2.46 1.05 Skn816S Skin 1 0.28 0.10 SmInt21XA Sm.Int. 1 6.43 12.04 Sto288S Stomach 1 7.41 14.32 Thr270T Thyroid 1 0.990.14 Tst647T Testis 1 1217.75 15.62 Utr135XO Uterus 1 237.21 55.14

[0341] In the analysis of matching samples, the level of mRNA expressionin cancer samples and the isogenic normal adjacent tissue from the sameindividual were compared. This comparison provides an indication ofspecificity for the cancer stage (e.g. higher levels of mRNA expressionin the cancer sample compared to the normal adjacent). Table 18 showsoverexpression of the LSG of SEQ ID NO:15 in 14 lung cancer tissuescompared with their respective normal adjacent (lung samples #1, 2, 3,4, 6, 7, 8, 10, 13, 14, 16,17,18, and 19). There is overexpression inthe cancer tissue for 70% of the lung matching samples tested (total of20 lung matching samples).

[0342] Altogether, the relative high level of lung tissue specificity,plus the mRNA overexpression in 70% of the lung carcinoma matchingsamples tested are believed to make the LSG of SEQ DI NO:15 a gooddiagnostic marker for lung cancer.

[0343] Primers used for expression analysis in this example are asfollows: Forward 5′ AAGGGAGCACCGTGGAGAA 3′ (SEQ ID NO:34) Reverse5′ AGGGCTGGATGACTTGGGA 3′ (SEQ ID NO:35) Probe5′ TTCCCAACTCTAACCCCACCCACG 3′ (SEQ ID NO:36)

[0344]

1 56 1 1449 DNA Homo sapien 1 ctgggagtgg atttcataca ttcgtggaggtggtgagagg atatactacg cagactctgt 60 gaggggccga ttcaccgtct ccagggacaacgccaagaac tcactctatc tgcaaatgaa 120 cagcctgaga gccgaggaca cggctgtttatttctgtgcg agagagccac cagcacccaa 180 ttactttgac tgctggagcc agggaaccctggtcaccgtc tcctcagctt ccaccaaggg 240 cccatcggtc ttccccctgg cgccctgctccaggagcacc tctgggggca cagcggccct 300 gggctgcctg gtcaaggact acttccccgaaccggtgacg gtgtcgtgga actcaggcgc 360 cctgaccagc ggcgtgcaca ccttcccggctgtcctacag tcctcaggac tctactccct 420 cagcagcgtg gtgaccgtgc cctccagcagcttgggcacc cagacctaca cctgcaacgt 480 gaatcacaag cccagcaaca ccaaggtggacaagagagtt gagctcaaaa ccccacttgg 540 tgacacaact cacacatgcc cacggtgcccagagcccaaa tcttgtgaca cacctccccc 600 atgcccacgg tgcccagagc ccaaatcttgtgacacacct cccccatgcc cacggtgccc 660 agcacctgaa ctcctgggag gaccgtcagtcttcctcttc cccccaaaac ccaaggatac 720 ccttatgatt tcccggaccc ctgaggtcacgtgcgtggtg gtggacgtga gccacgaaga 780 ccccgaggtc cagttcaagt ggtacgtggacggcgtggag gtgcataatg ccaagacaaa 840 gccgcgggag gagcagttca acagcacgttccgtgtggtc agcgtcctca ccgtcctgca 900 ccaggactgg ctgaacggca aggagtacaagtgcaaggtc tccaacaaag ccctcccagc 960 ccccatcgag aaaaccatct ccaaaaccaaaggacagccc cgagaaccac aggtgtacac 1020 cctgccccca tcccgggagg agatgaccaagaaccaggtc agcctgacct gcctggtcaa 1080 aggcttctac cccagcgaca tcgccgtggagtgggagagc agcgggcagc cggagaacaa 1140 ctacaacacc acgcctccca tgctggactccgacggctcc ttcttcctct acagcaagct 1200 caccgtggac aagagcaggt ggcagcaggggaacatcttc tcatgctccg tgatgcatga 1260 ggctctgcac aaccgcttca cgcagaagagcctctccctg tctccgggta aatgagtgcg 1320 acggccggca agcccccgct ccccgggctctcggggtcgc gcgaggatgc ttggcacgta 1380 ccccgtgtac atacttcccg ggcacccagcatggaaataa agcacccagc gctgccctgg 1440 gcccctgcg 1449 2 3825 DNA Homosapien misc_feature (428)..(428) a, c, g, or t 2 ttagtcacgt gaaaacccatcaggaaatta aacttgatga tagcaacatt cctcctccct 60 ctttaaaaac acgcccaccgtcaccaactt ttatcacaat agaatctact gcccgacgaa 120 cagaaaaccc tactaagaacgagctttctc agtcccctaa aaaggacagt tatgttgaac 180 ccccaccaag aaggcccatgtcgcaaaaat ctgaaattca cagagcaaac acttcccctt 240 ctccacccag gagtcgctctgaacaacttg tcagactcaa agacaccact gcaaagttat 300 ccaaaggggc catcccatgtccagcagcaa ccccggttcc aattgtagag aagaggtctg 360 aaatcatcat gtctcctgcaacacttcgtc gtcaaattaa gatagaaact cgtggtaggg 420 actctccnan ctacaatcacaataccagta aatataaatc atgctgctag tggttccttc 480 agagaatctg tggacgctcaagaggaaatc aggaaagtgg agaagagagc tacttatgtt 540 cataaagatg gactaaattccactgatcac atggtgcccg acactgaaag ttatgatgca 600 gttgaaatca tccgcaaggttgcagtgcct cctcgcctgt cagagcacac acagagatat 660 gaagcggcca accgaactgttcaaatggct gaaaatttcg tgaatgaccc tgaaaatgaa 720 ataaacagat ggttcagggaatttgagcat ggcccagttt ctgaagcaaa gtcaaataga 780 agagtttatg caaagggagaaacaaaccat aacatacaac aagaaagtcg tacattttgt 840 aaggaggaat ttggattaacatctttagga aacacgagtt ttacagactt ttcttgcaaa 900 catcctagag aactgcgagaaaagattcct gttaagcagc ccaggatctg ctctgaaacc 960 aggtctctaa gtgaacatttctcaggcatg gatgcatttg agagtcaaat tgttgagtcg 1020 aagatgaaaa cctcttcatcacatagctca gaagctggca aatctggctg tgacttcaag 1080 catgccccac caacctatgaggatgtcatt gctggacata ttttagatat ctctgattca 1140 cctaaagaag taagaaaaaattttcaaaag acgtggcaag agagtggaag agtttttaaa 1200 ggcctgggat atgcaaccgcagatgcttct gcaactgaga tgagaaccac cttccaagag 1260 gaatctgcat ttataagtgaagctgctgct ccaagacaag gaaatatgta tactttgtca 1320 aaagacagtt tatccaatggagtgcctagt ggcagacaag cagaattttc ataagtcctg 1380 cttccgatgc caccattgcaacagtaaact aagtttggga aattatgcat cacttcatgg 1440 acaaatatac tgtaaacctcactttaaaca acttttcaaa tccaaaggaa attatgatga 1500 aggttttgga cataagcagcataaagatag atggaactgc aaaaaccaaa gcagatcagt 1560 ggactttatt cctaatgaagaaccaaatat gtgtaaaaat attgcagaaa acacccttgt 1620 acctggagat cgtaatgaacatttagatgc tggtaacagt gaagggcaaa ggaatgattt 1680 gagaaaatta ggggaaaggggaaaattaaa agtcatttgg cctccttcca aggagatccc 1740 taagaaaacc ttaccctttgaggaagagct caaaatgagt aaacctaagt ggccacctga 1800 aatgacaacc ctgctatcccctgaatttaa aagtgaatct ctgctagaag atgttagaac 1860 tccagaaaat aaaggacaaagacaagatca ctttccattt ttgcagcctt atctacagtc 1920 cacccatgtt tgtcagaaagaggatgttat aggaatcaaa gaaatgaaaa tgcctgaagg 1980 aagaaaagat gaaaagnnggaaggaaggaa gaatgtgcaa gataggccga gtgaagctga 2040 agacacaaag agtaacaggaaaagtgctat ggatcttaat gacaacaata atgtgattgt 2100 gcagagtgct gaaaaggagaaaaatgaaaa aactaaccaa actaatggtg cagaagtttt 2160 acaggttact aacactgatgatgagatgat gccagaaaat cataaagaaa atttgaataa 2220 gaataataat aacaattatgtagcagtctc atatctgaat aattgcaggc agaagacatc 2280 tattttagaa tttcttgatctattaccctt gtcgagtgaa gcaaatgaca ctgcaaatga 2340 atatgaaatt gagaagttagaaaatacatc tagaatctca gagttacttg gtatatttga 2400 atctgaaaag acttattcgaggaatgtact agcaatggct ctgaagaaac agactgacag 2460 agcagctgct ggcagtcctgtgcagcctgc tccaaaacca agcctcagcc agaggcctta 2520 tggtaaaggg gggaagttcaatcatctctc ctgatacaaa tctcttaaac attaaaggaa 2580 gccattcaaa gagcaaaaatttacactttt tcttttctaa caccgtgaaa atcactgcat 2640 tttccaagaa aaatgagaacattttcaatt gtgatttaat agattctgta gatcaaatta 2700 aaaatatgcc atgcttggatttaagggaat ttggaaagga tgttaaacct tggcatgttg 2760 aaacaacaga agctgcccgcaataatgaaa acacaggttt tgatgctctg agccatgaat 2820 gtacagctaa gcctttgtttcccagagtgg aggtgcagtc agaacaactc acggtggaag 2880 agcagattaa aagaaacaggtgctacagtg acactgagta aaatatctat ggccactgac 2940 agtccacact taggcactgagagatattga tgttctgaaa taagatttta tgaatttgga 3000 tacccttttg aggaacttgatgtaaacatg gtgttcagaa atctcgtgtc tatctcaatg 3060 ggatatttct tgtattacaccttgtcattt ttttcacaat ttatttacat ctacttttgt 3120 ttgaactgga atgaagagatgaaacactat ggatatgttt tccattcaaa tggcacttta 3180 gcatattgtt ctgttttcctgtaaaacatc atgggtgtga tttttatact gctgctgctt 3240 gtcacaatta ttataacttctctgtaattt cctctgaaat aaaattgaat cacctgaggt 3300 gcaaaccaaa atacttctgtaacttttttt gatatatact gtcattctaa gtacatatac 3360 tccttgtgac ttgggaagtatttgtcttga ggcaagtatt taccacccac actaaaataa 3420 tgctggaaaa aataaaatactaaactgaag gcacagtatt attagaaagt gtaacatttt 3480 cattttctct tttactccacattttaaaga tacgagggtt attgttcttg aaataattac 3540 ctatattaaa ttatcatagaatgtgtctat aaacatttga cgaaaaatgt tgattttcct 3600 ccagaataat gtgaagtccatactcagaaa ttaactagaa aggttttaga cattacttaa 3660 ataaattatt cacattgcatttgtattgct tgctctgtgt aatggataag tataacaatc 3720 atatcactac agtttgtcaggttttcttct tatcatattt gatgaatatt aagtttttct 3780 gttatgaaaa catattcctctaaaatttgg cttctaaatt ttcta 3825 3 2315 DNA Homo sapien 3 gtaagcagcagttgattaga attaaatgag cttgaatttg attctgacat tcatattgat 60 ttgtccttccctcaaaaaac accctgagta tggacagggc ttcccgactc tgcagactac 120 acgccgtccatgagcagtgc ccaggtgtca ttacctgccc atgagatgtg acctgggcag 180 gggtccccacctgtaccctt gggccccagg agggaagccc agcatgtcag gctgaagcgg 240 gggtgcttccagagatggcc atgcagagca gccctcccgc ctcgggtcct gaggccccgc 300 tcagtggtccccccactctg cagaatgtgc acccccagct ctgatgtctc ttccaggtga 360 aatccgggtccccggccgtg ctggcattcg caaaggagaa gtcttttggg tggcccagct 420 tcatcacatacacggtcggc gtctcggacc ccgcggctgg cagccaaggg cctctgtcca 480 ctaccctgaccttctccagc cccgtgacca accaagccat tgccatccca gtgacagtgg 540 cttttgtgatggatcgccgt gggcccggtc cttatggagc cagcctcttc cagcacttcc 600 tggattcctaccaggtcatg ttcttcacgc tcttcgccct gttggctggg acagcggtca 660 tgatcatagcctaccacact gtctgcacgc cccgggatct tgctgtgcct gcagccctca 720 cgcctcgagccagccctggg acacagcccc cactatttcg ctgcctcatc acccacatct 780 cccaatgcattgcctcctgc tcgcaaagcc agccctccct cagggctgtg gagcccaggc 840 ctatggcctcccactagggc cgcgtgaagg ttcccggagg atggggtctc agccgagcct 900 cgttgcaacccccaagatgg aacatccctt gctgcattca cactggaaca agcccctcca 960 gatgagtgccccggccccag gccagcttca ctgccgtctc ttcacacaga gctgtagttt 1020 cggctctgcccattagctca ttttatgtag gagttttaaa tgtgtgtttt tttcctttca 1080 agtcttacaaagctaagact ttttggctca ttcctttttg catggttgtc tagggtttct 1140 ggacaatgtgctgttgcatt tttattttcc tagccttgct aaaatctttc ccttctcaag 1200 actttgagcagttagaagtg ctctttagaa gttgtctgtg ggtgatgtta ctgtagtggt 1260 ctcagggaaaggattgtcca gttactttag ggggtttttg gtggggtttt tccccctgtg 1320 aaaacttactttgcccctag tctggctgct gctaggactt ctgaggagca atgggacatg 1380 agtgtccctgtatctgcgcc actgccgcaa gggaagcctc aggaaccagc acctggaggc 1440 caggatagccaagccctggg tgagcgagag gctggagaac acaggagctc acccagggct 1500 gctgcccaaccatgggccac tgtgaacaga cttcagtcct ctgtttttgt ttcataagcc 1560 gttgagacatctgatggact tggcttaggc cctgctggga catcccacgt gtgatccctt 1620 tcactccatcaggacaccag gactgtcctt aggaaaatgt ccttgagatg gcagcaggag 1680 tcatattttctgtgtgtgtg tttcggaaag ccgctgtgtc ctgcctcagc acaaagaccc 1740 agtgtcatttgctcctcctg ttcctgtgcc actccagaac ctcagcagat ctgagccacc 1800 gcctgccagtgtgagaggcg gccactttca tggcagctca tcaggcgcag ggccccagac 1860 agcttcccagcaggccctag agcccggcct gggccaatga tggagggcgg ccgccagccc 1920 agggcctgcccatccagaag ggactcccca gggcctgggg gaggagaccc ttggaaaagt 1980 cctctcttcccagctcctga ttctggatct gagattctca gatcacaggc ccctgtgctc 2040 caggccgaggctgggctacc ctcagggaga tccagagact catgcccatg gccatccatg 2100 cgtggacgctgtgtggagag tccaggatga cgggatcccg cacaagctcc cttcagtcct 2160 tcagggctgggccatgtggt tgatttttct aaagctggag aaaggaagaa ttgtgccttg 2220 catattacttgagcttaaac tgacaacctg gatgtaaata ggagcctttc tactggttta 2280 tttaataaagttctatgtga tttttaaaaa aaaaa 2315 4 300 DNA Homo sapien misc_feature(8)..(8) a, c, g or t 4 agcatganaa aggtgaaggc tgccggtggc acggggctcggatctgctgc cgggccgacc 60 tgggagagcc atgaggctgt atgtgatggg gcacctcttgggtgcacact ttggatgaca 120 agtgccccca agaggagcca gggctggctg cagtgaggccccaggaggtt ctccaggggc 180 gtcctgcttc agctcaaggg gctaggaata ggggaaacgatgcagggaag ccaatggccc 240 aagtggctcc ctcactgact gttacttgct gtgtatgtctctttcttttc ttttttttcc 300 5 4347 DNA Homo sapien 5 gcggtgcggc ggcgggaggcggaggcgagg gtgcgatggc gcggagcccg ggacgcgcgt 60 acgccctgct gcttctcctgatctgcttta acgttggaag tggacttcac ttacaggtct 120 taagcacaag aaatgaaaataagctgcttc ctaaacatcc tcatttagtg cggcaaaagc 180 gcgcctggat caccgcccccgtggctcttc gggagggaga ggatctgtcc aagaagaatc 240 caattgccaa gatacattctgatcttgcag aagaaagagg actcaaaatt acttacaaat 300 acactggaaa agggattacagagccacctt ttggtatatt tgtctttaac aaagatactg 360 gagaactgaa tgttaccagcattcttgatc gagaagaaac accatttttt ctgctaacag 420 gttacgcttt ggatgcaagaggaaacaatg tagagaaacc cttagagcta cgcattaagg 480 ttcttgatat caatgacaacgaaccagtgt tcacacagga tgtctttgtt gggtctgttg 540 aagagttgag tgcagcacatactcttgtga tgaaaatcaa tgcaacagat gcagatgagc 600 ccaataccct gaattcgaaaatttcctata gaatcgtatc tctggagcct gcttatcctc 660 cagtgttcta cctaaataaagatacaggag agatttatac aaccagtgtt accttggaca 720 gagaggaaca cagcagctacactttgacag tagaagcaag agatggcaat ggagaagtta 780 cagacaaacc tgtaaaacaagctcaagttc agattcgtat tttggatgtc aatgacaata 840 tacctgtagt agaaaataaagtgcttgaag ggatggttga agaaaatcaa gtcaacgtag 900 aagttacgcg cataaaagtgttcgatgcag atgaaatagg ttctgataat tggctggcaa 960 attttacatt tgcatcaggaaatgaaggag gttatttcca catagaaaca gatgctcaaa 1020 ctaacgaagg aattgtgacccttattaagg aagtagatta tgaagaaatg aagaatcttg 1080 acttcagtgt tattgtcgctaataaagcag cttttcacaa gtcgattagg agtaaataca 1140 agcctacacc cattcccatcaaggtcaaag tgaaaaatgt gaaagaaggc attcatttta 1200 aaagcagcgt catctcaatttatgttagcg agagcatgga tagatcaagc aaaggccaaa 1260 taattggaaa ttttcaagcttttgatgagg acactggact accagcccat gcaagatatg 1320 taaaattaga agatagagataattggatct ctgtggattc tgtcacatct gaaattaaac 1380 ttgcaaaact tcctgattttgaatctagat atgttcaaaa tggcacatac actgtaaaga 1440 ttgtggccat atcagaagattatcctagaa aaaccatcac tggcacagtc cttatcaatg 1500 ttgaagacat caacgacaactgtcccacac tgatagagcc tgtgcagaca atctgtcacg 1560 atgcagagta tgtgaatgttactgcagagg acctggatgg acacccaaac agtggccctt 1620 tcagtttctc cgtcattgacaaaccacctg gcatggcaga aaaatggaaa atagcacgcc 1680 aagaaagtac cagtgtgctgctgcaacaaa gtgagaaaaa gcttgggaga agtgaaattc 1740 agttcctgat ttcagacaatcagggtttta gttgtcctga aaagcaggtc cttacactca 1800 cagtttgtga gtgtctgcatggcagcggct gcagggaagc acagcatgac tcctatgtgg 1860 gcctgggacc cgcagcaattgcgctcatga ttttggcctt tctgctcctg ctattggtac 1920 cacttttact gctgatgtgccattgcggaa agggcgccaa aggctttacc cccatacctg 1980 gcaccataga gatgctgcatccttggaata atgaaggagc accacctgaa gacaaggtgg 2040 tgccatcatt tctgccagtggatcaagggg gcagtctagt aggaagaaat ggagtaggag 2100 gtatggccaa ggaagccacgatgaaaggaa gtagctctgc ttccattgtc aaagggcaac 2160 atgagatgtc cgagatggatggaaggtggg aagaacacag aagcctgctt tctggtagag 2220 ctacccagtt tacaggggccacaggcgcta tcatgaccac tgaaaccacg aagaccgcaa 2280 gggccacagg ggcttccagagacatggccg gagctcaggc agctgctgtt gcactgaacg 2340 aagaattctt aagaaattatttcactgata aagcggcctc ttacactgag gaagatgaaa 2400 atcacacagc caaagattgccttctggttt attctcagga agaaactgaa tcgctgaatg 2460 cttctattgg ttgttgcagttttattgaag gagagctaga tgaccgcttc ttagatgatt 2520 tgggacttaa attcaagacactagctgaag tttgcctggg tcaaaaaata gatataaata 2580 aggaaattga gcagagacaaaaacctgcca cagaaacaag tatgaacaca gcttcacatt 2640 cactctgtga gcaaactatggttaattcag agaataccta ctcctctggc agtagcttcc 2700 cagttccaaa atctttgcaagaagccaatg cagagaaagt aactcaggaa atagtcactg 2760 aaagatctgt gtcttctaggcaggcgcaaa aggtagctac acctcttcct gacccaatgg 2820 cttctagaaa tgtgatagcaacagaaactt cctatgtcac agggtccact atgccaccaa 2880 ccactgtgat cctgggtcctagccagccac agagccttat tgtgacagag agggtgtatg 2940 ctccagcttc taccttggtagatcagcctt atgctaatga aggtacagtt gtggtcactg 3000 aaagagtaat acagcctcatgggggtggat cgaatcctct ggaaggcact cagcatcttc 3060 aagatgtacc ttacgtcatggtgagggaaa gagagagctt ccttgccccc agctcaggtg 3120 tgcagcctac tctggccatgcctaatatag cagtaggaca gaatgtgaca gtgacagaaa 3180 gagttctagc acctgcttccactctgcaat ccagttacca gattcccact gaaaattcta 3240 tgacggctag gaacaccacggtgtctggag ctggagtccc tggccctctg ccagattttg 3300 gtttagagga atctggtcattctaattcta ccataaccac atcttccacc agagtcacca 3360 agcatagcac tgtacagcattcttactcct aaacagcagt cagccacaaa ctgacccaga 3420 gtttaattag cagtgactaatttcatgttt ccaatgtacc tgatttttca tgagccttac 3480 agacacacag agacacatacacattgatct taaaattttt ctcagtcact gatatgcaaa 3540 ggaccacact gtctctgcttccaggagtat tttagaaatg ttccacaatt tactgaagac 3600 atagagatga tgctgctgcttaggtgcctt ttagcaagct atgcaaacaa tcctgataaa 3660 acaagataca tagagagtcaatctggcttc tgagaattta ccaagtgaac agagtaccta 3720 gttcatcagc cgtccagtaaagcaacccag gaaactgact gggtctcttt gcctaccgta 3780 ttaacattaa acattgatgttctgtattct gtactttact gcacccagca gactttcaac 3840 aactcattga cccaaagtgctgggattaca ggcgtgagcc actgcgcccg gccacattca 3900 gttcttatca aagaaataacccagacttaa tcttgaatga tacgattatg cccaatatta 3960 agtaaaaaat ataagaaaaggttatcttaa atagatctta ggcaaaatac cagctgatga 4020 aggcatctga tgccttcatctgttcagtca tctccaaaaa cagtaaaaat aaccactttt 4080 tgttgggcaa tatgaaatttttaaaggagt agaataccaa atgatagaaa cagactgcct 4140 gaattgagaa ttttgatttcttaaagtgtg tttctttcta aattgctgtt ccttaatttg 4200 attaatttaa ttcatgtattatgattaaat ctgaggcaga tgagcttaca agtattgaaa 4260 taattactaa ttaatcacaaatgtgaagtt atgcatgatg taaaaaatac aaacattcta 4320 attaaaggct ttgcaacacaaaaaaaa 4347 6 2116 DNA Homo sapien 6 tagcgacctc tcgcagggaa agtcagcgtcggccaaaagc ctccgggatc ggaatgagga 60 ggctgctgga gaagttgctt tctcctaaaagggattatcc caggcgcaca cggtcattac 120 acgccgggga cgctcagtcg gtgcgggtacccctgggcag ggccagcccc gcattccagg 180 ttctccatgt gcctagaaga cagtaatcgacggtatagca acagatctga ctgctaacat 240 gcgaaaccga tcagtagtag cagtagcaccagcaacagca gcacgaaaag caaaactaat 300 ctaaacggcc ctcagggtct aagcaggcccgacgaagact cgcccatccg gtcgccagaa 360 aactgggagt cccgcccgcc tttccggcactgaaacgcga tcggccctgc ctggtaccgc 420 atcctcctct tgaccccacc tacactacgacgacggacgc tcgagaacgc ggcaccgcgc 480 cccgcaggaa gtgcttccct gggcggaagcttctgagcgt gatatagcgg aagtgccttc 540 tcttccggtc tttctggtct cggccgcagaagcgagatga cgaagggaac gtcatcgttt 600 ggaaagcgtc gcaataagac gcacacgttgtgccgccgct gtggctctaa ggcctaccac 660 cttcagaagt cgacctgtgg caaatgtggctaccctgcca agcgcaagag aaagtataac 720 tggagtgcca aggctaaaag acgaaataccaccggaactg gtcgaatgag gcacctaaaa 780 attgtatacc gcagattcag gcatggattccgtgaaggaa caacacctaa acccaagagg 840 gcagctgttg cagcatccag ttcatcttaagaatgtcaac gattagtcat gcaataaatg 900 ttctggtttt aaaaaataca tatctggttttggtaaggta tttttaatca attaggcttg 960 tagtatcagt gaaatactgt aggtttagggactgggctag cttcatatca gatttacttg 1020 ttaagtgact gttttggaat gtttacttttggactgggtt tgtaacacgg ttaaaggcaa 1080 tgagaaacaa gcagaattcc aggagtccttgaagcagagg gcactggaag acaatatagc 1140 agattaaaat agcacagctc atgtggcataggtgggtatt ttagatgttt gagtaaattt 1200 gaaagagtat gatgtttaaa ttacctttagcaacatgttc atctgctatg ctgtcatgac 1260 tagggggatg attattagtc acatagagcttgggagtacc actggaaacg tatgggtagg 1320 agtttaggtg gcttctgttt ttcaaaagatgatcttatcc tagtatctgt aatgctcact 1380 tggcacacct gacttgtggg ctgtgtgtaaggtggctagc taagtgaaaa aagcctgcta 1440 ggtgtgagtc aacttaagaa tatgtaaataggtttgagaa aaagtagggc ttgggtgcaa 1500 gtaaagattg agcaggaaat aaaggaaaatcaagtataat ccctgagatt tgtagattaa 1560 aggcaatgat gtgggactac ttggtcgaatttttttagcc ctcaacttgg taattgggtg 1620 tttctgtgtt aaagcactga aacttgctgtcgtgccttcc tagttttcgt ggtttattga 1680 cagggttggg ggtttttttt gtttttttaaaatgaaggga caaagtcaac tggactgctg 1740 agtgagaggg caggggcagt tgaagggaacatgaattgct ggaacagcta cataaaatag 1800 tgatgtagcc aagtcatgct atttaaattataattctcca ctgtgtttag aataacatct 1860 gaggttctta acctggcctt ggaagggtatcacttttact tgtaacctgg aatggcttta 1920 taatgtgcta gctaattgct actctcatcttgtattttaa ctcctaattt acccttcagg 1980 tctcagcttc agaacattca cttataaagaaaccctgctg attaaatctc tcttgggctt 2040 cctcccgaaa tgtgagacta tactttaaagatgtatggtt agagtccaat tgccattgcc 2100 tttcttgttt acagat 2116 7 4474 DNAHomo sapien misc_feature (9)..(9) a, c, g or t 7 cggcccggnc gggggggcaagatggcggcg gcagtagggg ttcgtggccg gtacgagctg 60 ccgccttgct ccggcccaggctggctcctc agcctttccg ccttgctgag tgtggcggca 120 cgaggggcct tcgccaccacgcactgggtc gtcacggagg acgggaaaat ccagcagcag 180 gtggattcac caatgaacttgaagcatcct catgacctag tcatattaat gagacaagaa 240 gcaacagtta actacctcaaagaattagag aaacaattag ttgctcaaaa aattcacata 300 gaagagaatg aggacagagacacaggactg gaacagagac ataataaaga agacccagac 360 tgcatcaaag ccaaggtgcccttaggggac ctggatctat atgatggcac atacataact 420 ttggagagca aagacatcagtcctgaagat tatatagaca cagaatctcc tgtccctcca 480 gacccagagc aacctgattgtactaaaatt ctagaacttc catatagtat acatgctttt 540 cagcacttga gaggtgtacaggagagagtt aatctttctg cacctctgct acctaaagaa 600 gacccaatct tcacatatttatctaaacgg ttaggaagga gtatagatga cataggtcac 660 ctcattcatg aaggcctacagaagaacact tcctcgtggg tactgtataa catggcttca 720 ttttactgga gaattaagaatgagccatat caggtagtag aatgtgccat gcgagcactt 780 cacttctctt ccaggcacaataaagacatt gccctggtca acctggcaaa cgttctacac 840 agagcacact tctctgctgatgctgctgtc gtggtccatg cagctctgga tgacagtgac 900 ttcttcacca gctattacactttggggaat atatatgcaa tgcttgggga atataaccac 960 tcagtgctct gttatgaccacgctttgcag gccagacctg ggtttgagca agctataaag 1020 aggaagcatg ctgtcctatgtcagcaaaaa ctggagcaga aattggaggc tcagcataga 1080 tctctccagc gaacactgaatgagttaaaa gagtatcaaa agcagcatga ccactacctg 1140 agacagcagg aaatcctagaaaaacataaa ctgattcagg aggagcaaat cttaagaaat 1200 atcattcatg agactcagatggcaaaagag gcacaattag gaaatcatca gatatgccga 1260 ctggtcaacc agcagcatagtttacattgc cagtgggacc agcctgtacg ctatcatcgt 1320 ggagatatct ttgaaaatgtggactatgtt cagtttggtg aggattcatc aacctccagt 1380 atgatgtctg tgaactttgatgttcaatca aatcagagtg atatcaatga ttcggtcaag 1440 tcttctcccg tagcccattctattctctgg atttggggca gggactctga tgcatatagg 1500 gacaaacagc atattctatggcctaaaaga gcagattgta cagaaagcta ccctagagtc 1560 cctgttggtg gggaattgccaacgtatttt ctgcctccgg aaaacaaagg actcaggatc 1620 cacgaactca gcagtgatgattattctaca gaagaagagg cccaaacccc tgactgttcc 1680 ataactgact tcagaaaaagccacactctg tcctacttag tcaaagaatt agaggttcgc 1740 atggatctga aagccaaaatgccagatgac catgcacgaa aaattttgct ttcccgtatt 1800 aataactata ctatcccagaagaagaaatt gggtctttct tatttcatgc tattaataag 1860 ccaaatgctc ctatctggctcatactcaat gaagctggac tatactggag agcagtagga 1920 aatagcactt ttgctattgcctgtcttcag agggctttga atttagctcc acttcaatac 1980 caagatgttc ctcttgtcaacttggccaac cttttgattc attacggcct tcatcttgat 2040 gccactaagc tgctacttcaagctttggcc atcaatagct ctgagcctct gacctttttg 2100 agcctgggaa atgcttaccttgctctgaag aatatcagtg gggcacttga ggcctttaga 2160 caggccttga aattaaccaccaaatgtcca gagtgtgaaa acagcctgaa gttgatccgc 2220 tgtatgcagt tttatccttttctgtacaac atcacttctt ctgtttgcag tggtacggtg 2280 gttgaggaga gcaatggttctgatgagatg gagaattcag atgaaaccaa aatgtcagaa 2340 gaaatactgg ctttggtggatgaatttcaa caggcatggc ctttggaagg ctttgggggt 2400 gcactagaga tgaaagggcggcgtctagac ttacaaggaa tacgggtgct gaagaaaggt 2460 ccccaggatg gagtggccagaagctcttgc tatggagact gcagaagtga agatgatgaa 2520 gcaacagaat ggattacattccaggtcaaa cgtgtaaaga aacccaaagg agatcataag 2580 aaaactcctg ggaaaaaagtagaaacaggt cagatagaaa atggacatcg ttaccaagca 2640 aacctagaga tcactggccccaaggtggca tctcctgggc cacaaggaaa aaaacgtgac 2700 taccagcgtc tgggatggcccagcccggac gaatgcctca aactccgctg ggtagagctg 2760 actgccatcg tgagtacctggcttgcagtt tcttcaaaaa acattgacat cacagaacac 2820 atagattttg ccacccctatacagcagcca gcaatggagc ctctttgcaa tggcaatctc 2880 cccacgagta tgcataccctggaccacttg catggggttt ccaaccgagc cagcctgcac 2940 tacacagggg agagtcagttaacagaggta ttacaaaatc tcggcaaaga ccaatatcca 3000 caacagtcgc ttgaacagattggcacccga attgccaaag ttttggaaaa gaaccagacg 3060 tcctgggtcc tctccagcatggcagccctc tactggaggg tgaaaggcca aggaaagaag 3120 gcaatcgact gcctccgccaggctctgcac tatgcgccac accagatgaa ggatgtgccc 3180 ctgattagcc tggccaacatcttgcacaat gccaagctct ggaatgacgc cgtcatagta 3240 gccaccatgg cagtagagatcgcaccacac tttgctgtga accacttcac tctgggcaat 3300 gtctacgtgg caatggaagaatttgaaaaa gcactggtgt ggtatgaatc cacattgaag 3360 cttcagcccg agtttgtcccagccaagaac cgaatccaga ccatccagtg tcacttaatg 3420 ctgaagaagg gacggcgctctccttagtgc acttcttcct tctctctttc tctttactca 3480 tgctctaaaa aaaaagaataagaaaagaaa ccaatcattg tcagtatcta ctattaatga 3540 tgtgtgtgaa aataactaagacttataaca ggacttttac atatgtggga attggtttgt 3600 ttttgttttt acgtttctcctttcccccaa ccaacctcag aagaggcacc ttcagaaaca 3660 cacatttctt aaaaggaaagtgcagcttca agatattgtg taaatactga gccaagacat 3720 ttctggagct gtgctctgtctccaaaaacc tcaatgcctt tagggctttt ctcagtggtc 3780 cagctagcct tctctttggaggaggatgaa gccgcattgc acattctctg cttcctgtcg 3840 tagcctctgt tgtcaatggaaatgcggaag cccatctggt gcccgtcagt gagaagcaac 3900 gttctgcgct ctctccggtagacctccatg ctgtccccag tcttgtccat tccatgctgc 3960 tgtgttacaa actctcagaggtagtttgca ggggaggaag gggaatatga ttttaaaaac 4020 aaaatattta caacaacaaaaattcttagg atcacctgac ctttgtaatg ttatttatgt 4080 tggggaggga ggggggctgagaaggggaaa tcagcagtgt gcaacatctt tataatttgt 4140 actttaatta caaatcacaaggaaaccaat aagttgaaat cctatataac aggtttatat 4200 atatagaata tgtatatttgaagccctcta cagactgagt ctatgtttta ctaattcttt 4260 gttcactgtg ttacccatcttggaataagt tgtgaatgtc agctccctct ctctgaggcc 4320 tccagactta gctcctcaggagggtaatga gccaaggttg agtgtttcca tacaatgctt 4380 ttacctttga tcccaggagaatcagaaact ccaacatttt ggaatcttca agggcacata 4440 ctgagaaaaa aaataaaattgtttatgagc aaaa 4474 8 777 DNA Homo sapien misc_feature (269)..(439) a,c, g or t 8 aaataaataa ataaaattta gattaatttg ctgttatatt tttatataaactatgacata 60 agtataaaca aaaaaataga ataagtaaat aaataaataa aatttagattaatttgctgt 120 tacattttta tataagctat gtttatgaca gactttccta taatattcttatcataatgt 180 tcttgcactt gaaagaatgt gcattctgca gttgtgtgca ggtgttatgtgtatttcaac 240 tgggtcaagt ttgttaatca ggttggtcnn nnnnnnnnnn nnnnnnnnnnnnnnnnnnnn 300 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnnnnnnnnnnnn 360 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnnnnnnnnnnnn 420 nnnnnnnnnn nnnnnnnnna cgttttctgg tagagtgaac cctttattatgtgaaatggc 480 tttctctacc actcacagtg gtattttttt cactttcccc cttatttttgaattttcagt 540 gttcttataa tgatgtcatg tctcttgaat agagttgttt taatcaagtctgtcaatttt 600 tgtcttttgt gtatttcgtg aatttacatt tattgtaatt actgatatagttgtgttcat 660 aaataccatc ttactatttg tttcccattt gtcttaccca tttttgtttttattnntttc 720 ttttctgttg cctttccttt ccngattaan ttnttaattc ccgttttccccccccta 777 9 3195 DNA Homo sapien 9 ctctctttta gtgtcactgt caatggcgctacatggactt tgtaataacc ctttgaggca 60 catagctggg tgccatgtag aacatgtatctgttacgata agtgtgtgcc caagaaatca 120 gaagaatgga ctttaatctc attttagaaagtatgatatt aaatgattta cccaagccat 180 acttcaggtt aatgacacga tagaagctagtacctgtgtc tctcaaattt ttctaacact 240 ttttatcttc cgatcaggtt tgtgcaggggaaactcagtg gagaggaaga tatatatccc 300 cttaaataaa acagctccct gtgttcgcctgctcaacgcc actcatcaga ttggctgcca 360 gtcttcaatt agtggagaca caggggttatccacgtagta gagaaagagg aggacctaca 420 gtgggtattg actgatggcc ccaaccccccttacatggtt ctgctggaga gcaagcattt 480 taccagggat ttaatggaga agctgaaagggagaaccagc cgaattgctg gtcttgcagt 540 gtccttgacc aagcccagtc ctgcctcaggcttctctcct agtgtacagt gcccaaatga 600 tgggtttggt gtttactcca attcctatgggccagagttt gctcactgca gagaaataca 660 gtggaattcg ctgggcaatg gtttggcttatgaagacttt agtttcccca tctttcttct 720 tgaagatgaa aatgaaacca aagtcatcaagcagtgctat caagatcaca acctgagtca 780 gaatggctca gcaccaacct tcccactatgtgccatgcag ctcttttcac acatgcatgc 840 tgtcatcagc actgccacct gcatgcggcgcagctccatc caaagcacct tcagcatcaa 900 cccagaaatc gtctgtgacc ccctgtctgattacaatgtg tggagcatgc taaagcctat 960 aaatacaact gggacattaa agcctgacgacagggttgtg gttgctgcca cccggctgga 1020 tagtcgttcc tttttctgga atgtggccccaggggctgaa agcgcagtgg cttcctttgt 1080 cacccagctg gctgctgctg aagctttgcaaaaggcacct gatgtgacca ccctgccccg 1140 caatgtcatg tttgtcttct ttcaaggggaaacttttgac tacattggca gctcgaggat 1200 ggtctacgat atggagaagg gcaagtttcccgtgcagtta gagaatgttg actcatttgt 1260 ggagctggga caggtggcct taagaacttcattagagctt tggatgcaca cagatcctgt 1320 ttctcagaaa aatgagtctg tacggaaccaggtggaggat ctcctggcca cattggagaa 1380 gagtggtgct ggtgtccctg ctgtcatcctcaggaggcca aatcagtccc agcctctccc 1440 accatcttcc ctgcagcgat ttcttcgagctcgaaacatc tctggcgttg ttctggctga 1500 ccactctggt gccttccata acaaatattaccagagtatt tacgacactg ctgagaacat 1560 taatgtgagc tatcccgaat ggctgagccctgaagaggac ctgaactttg taacagacac 1620 tgccaaggcc ctggcagatg tggccacggtgctgggacgt gctctgtatg agcttgcagg 1680 aggaaccaac ttcagcgaca cagttcaggctgatccccaa acggttaccc gcctgctcta 1740 tgggttcctg attaaagcca acaactcatggttccagtct atcctcaggc aggacctaag 1800 gtcctacttg ggtgacgggc ctcttcaacattacatcgct gtctccagcc ccaccaacac 1860 cacttatgtt gtacagtatg ccttggcaaatttgactggc acagtggtca acctcacccg 1920 agagcagtgc caggatccaa gtaaagtcccaagtgaaaac aaggatctgt atgagtactc 1980 atgggtccag ggccctttgc attctaatgagacggaccga ctcccccggt gtgtgcgttc 2040 tactgcacga ttagccaggg ccttgtctcctgcctttgaa ctgagtcagt ggagctctac 2100 tgaatactct acatggactg agagccgctggaaagatatc cgtgcccgga tatttctcat 2160 cgccagcaaa gagcttgagt tgatcaccctgacagtgggc ttcggcatcc tcatcttctc 2220 cctcatcgtc acctactgca tcaatgccaaagctgatgtc cttttcattg ctccccggga 2280 gccaggagct gtgtcatact gaggaggaccccagcttttc ttgccagctc agcagttcac 2340 ttcctagagc atctgtccca ctgggacacaaccactaatt tgtcactgga acctccctgg 2400 gcctgtctca gattgggatt aacataaaagagtggaacta tccaaaagag acagggagaa 2460 ataaataaat tgcctccctt cctccgctcccctttcccat caccccttcc ccatttcctc 2520 ttccttctct actcatgcca gattttgggattacaaatag aagcttcttg ctcctgttta 2580 actccctagt tacccaccct aatttgcccttcaggaccct tctacttttt ccttcctgcc 2640 ctgtacctct ctctgctcct cacccccacccctgtaccca gccaccttcc tgactgggaa 2700 ggacataaaa ggtttaatgt cagggtcaaactacattgag cccctgagga caggggcatc 2760 tctgggctga gcctactgtc tccttcccactgtcctttct ccaggccctc agatggcaca 2820 ttagggtggg cgtgctgcgg gtgggtatcccacctccagc ccacagtgct cagttgtact 2880 ttttattaag ctgtaatatc tatttttgtttttgtctttt tcctttattc tttttgtaaa 2940 tatatatata atgagtttca ttaaaatagattatcccaca cgacttgtac tgctagttat 3000 tcttcccagg ccaccttgtt cagcgagcctagactggaag tcatgaagtt atcttttatg 3060 ctatcatctt gggctccaga ggacccaaggagtaaggctc tgtcaaaaac agttgaagtc 3120 ctttcaaatt gcagagcctg gttctcctcttgtaagaaca atgttaacat agtttcttct 3180 cactttgtaa tgaac 3195 10 949 DNAHomo sapien 10 agcctccacc ctggcgatgg ctccctggtc ctactttctc tctcaaactggctttttctc 60 attcctttga ctccgccaga cttcctcgcc cccatgacct ggtgttgtgtctgatcaccc 120 caacattcct ggctgcccaa tgtggggcaa tgaagacccc agtgaaggaatgctagagtg 180 tgtgaaagtg gaggacgcat cgtcaaagga cacctgagga cgtctcaaagaagctcggcg 240 ggagagctga gcgctcggaa gaaccaagaa tcatctcttt tgaaaaatcgattcatcaaa 300 tgaatcttca gccaacaact gttcaagaag gattcaaata tcacaggttccaagaagtaa 360 agctttggag gtcacaaaat tagcaataga agctgggttc cgccatatagattctgctca 420 tttatacaat aatgaggagc aggttggact ggccatccga agcaagattgcagatggcag 480 tgtgaagaga gaagacatat tctacacttc aaagcttggt ccacttttcatcgaccagag 540 ttggtccgac cagccttgga aaactcactg aaaaaagctc aattggactatgttgacctc 600 tatcttattc attctccaat gtctctaaag ccaggtgagg aactttcaccaacagatgaa 660 caagtggcaa aagtaatatt tgacatagtg gatctctgta ccaccctgggagggcatgga 720 gaaagtgtaa ggatggcagg aattggggca agtccattgg ggtgtcacacttcaacccgc 780 aggcgctggg gatgagtctc aaaaagcagg aatccagtta aagcggtctgcacccgtgga 840 gtgtccgatt taccgggtaa tgctgattcg gcagccagaa atgttggtgcaaagctgggg 900 ccccacaaat ggggcccccc ccgctgtggg ccctggttga aaaaccctg 94911 14917 DNA Homo sapien 11 gctcgagatc cattgtgctc taaaggtgaa aagacctagatggtagaagg caagcagtgg 60 aatgtgtctg ggtatccgag aaatatacag aaagcacttaagagaactta atcatttttt 120 tctccctttc cttagattga atagggaaaa cctgctttctgcaaacaact gaaaaagctg 180 catttagaaa ctgcttcttt ggccctcatc gagaagctggaacttgaatt gttaggcccc 240 ttatgggaca agctctcaac tgctgatcac ccagtgattgacaccatggc cagcaagagg 300 aaatccacca caccatgcat gatcccagtg aagactgtggtgttgcaaga tgccagcatg 360 gaggcccagc ccgctgagac cttgcctgaa ggaccccagcaggatctgcc cccagaagca 420 tctgctgcca gcagtgaggc agcacagaac cccagcagtactgatggctc tacactggcc 480 aatgggcatc ggagcacttt agatggctat ttatattcctgtaaatactg cgatttcaga 540 tcccatgaca tgacccaatt tgtgggacat atgaactcagagcacacaga ctttaataaa 600 gacccaacct ttgtatgcag tgggtgcagt tttctggcaaaaacccctga ggggctttcc 660 ttgcacaatg ccacatgtca ctccggggaa gccagctttgtgtggaacgt ggccaagcca 720 gacaatcatg tggttgtgga gcagagcatc cctgagagcaccagcactcc tgacctagcg 780 ggtgagccca gtgctgaagg ggctgatgga caggcagaaatcatcattac caaaactcca 840 atcatgaaga taatgaaagg caaagctgaa gccaaaaaaattcatacact caaggagaat 900 gtccctagcc agcctgtggg tgaggcctta ccaaagctgtcgactggaga aatggaggtg 960 agagaggggg accattcctt catcaatggg gcagttccagtcagccaggc atctgccagc 1020 tctgcaaaaa acccccatgc cgccaacggg cccctgataggaacagtgcc agttttgcca 1080 gctggcatag cacagttcct ctccctccag cagcagcccccagtgcatgc ccaacaccat 1140 gtccaccagc cactgcccac ggccaaggcc cttcccaaagtgatgatccc cctgagcagc 1200 attccaacgt acaatgcagc catggactct aacagcttcctgaagaactc cttccacaag 1260 ttcccctacc ccaccaaagc cgagctctgc tatttgactgtggtgaccaa gtatccagaa 1320 gaacagctca agatctggtt cacagcccaa aggctgaagcaggggatcag ctggtcccct 1380 gaggagattg aggatgcccg gaaaaagatg ttcaatacagtcatccagtc tgtgcctcag 1440 cccacaatta cggttctaaa taccccactc gtcgccagtgctggcaatgt ccagcatctc 1500 atccaggccg ctcttccagg tcacgttgtg gggcagccagagggtacagg agggggactt 1560 ctggtcactc agccattgat ggccaatggg ttgcaagcaacaagttcccc tctccccctc 1620 acggtgacat ccgtccccaa gcagccaggt gtggcacccattaacactgt gtgttcaaat 1680 acaacgtcag ctgtgaaggt ggtcaatgcg gcccagtcgctcctcacggc ctgccccagc 1740 ataacctccc aagccttcct tgatgctagc atctacaaaaataagaaatc tcatgaacag 1800 ctgtcagctc tgaaagggag cttctgtcgg aaccagttcccagggcagag cgaagttgaa 1860 catctcacaa aagtgacggg cctcagtacc agagaggtgcggaaatggtt cagtgatcgt 1920 agataccact gccggaactt gaagggctcc agagcgatgatacctggaga tcacagttcc 1980 atcatcattg actctgtgcc agaggtgtcc ttctccccatcgtccaaggt ccctgaggta 2040 acctgcattc cgacaacagc cacactagca acccacccttctgccaaacg acaatcttgg 2100 caccagactc ctgacttcac accaaccaaa tacaaggagagagcccctga gcagctcaga 2160 gccctggaga gcagttttgc acaaaaccct cttcctcttgatgaggaact ggaccgcctg 2220 agaagtgaaa ccaaaatgac ccgacgagaa attgatagctggttttcaga gagacggaaa 2280 aaagtgaatg ctgaggagac caagaaggct gaggagaatgcctctcagga ggaagaggag 2340 gctgctgagg atgagggtgg agaagaggat ttggccagtgagctaagggt ctctggtgaa 2400 aatggctctc tggaaatgcc cagcagccat atcttggcagagcgcaaagt cagccccatt 2460 aaaatcaacc tgaagaacct gagggtcact gaagccaatggcaggaacga gattccaggg 2520 ctgggtgcct gtgaccctga ggatgatgag tcaaacaaactggcagagca gctcccaggc 2580 aaagtgagct gcaaaaagac tgcccagcag cggcacttgctgcggcagct ctttgtccag 2640 acacagtggc caagcaacca ggactatgac tccatcatggcccagacggg tctgccacgg 2700 ccagaggtgg tgcgctggtt tggagatagc aggtacgcactgaagaacgg ccaactcaaa 2760 tggtacgaag actataagcg aggcaacttc ccaccagggctactggtcat tgcccctggc 2820 aaccgggagc tcctgcagga ctattacatg acacacaagatgctgtatga agaggacctg 2880 cagaacctct gtgacaagac ccagatgagc tcccagcaggtcaagcagtg gtttgctgag 2940 aaaatggggg aggagaccag agccgtggca gacacaggcagtgaggacca gggccctggt 3000 actggtgagc tcacagcagt tcacaaaggg atgggtgacacctattcaga ggtgtctgag 3060 aacagtgagt cgtgggagcc ccgtgtccct gaggccagctcagagccctt tgacacatcg 3120 agtccccagg ctggacgtca gctcgaaaca gactgaatttgatctgatta atgtgaagga 3180 ctggccagtc tgggaaaccg cctgccacgt ggaagagccaaacccgactc tctgctgcca 3240 catgccgttc ccatgcccgg ctgctgggca cctgggagagcttccagaat cctcgcagac 3300 agcccagagc ctgccgctac cctcggcctg cccaccaccaagcaagcagc aagcaagatg 3360 gggttctcat cagttcttcc tcccacaatg taggacctttcctttacctt ccaatggata 3420 aaatagttca gagttcatag tcatattcat agacacagaatcaagctttt aacatataca 3480 tccacctcta tatgttaaat aaaacatcag attatcaacactgtcattac gtagaaactt 3540 tggttagcca agcagtgcat tgtcagttac gtcatctctaaaaatgacct gtgtctgttc 3600 tctggggatt gctgggtcac aggtgcccct caccttccacagtcaggcag ggaagttata 3660 ggcacaaagc tacgtctgga acccctttgt gccccctttgtgttcctcaa ggaagcagta 3720 cctttgaaga gatctctgct gcattaagtg atgaccggctacgtttcatg tcaggcttgc 3780 tttgccttgt gggctactca gtgcagaacc tgctgtaaccctcagttcaa aaaatggact 3840 ggcaatgtga ttagcgttgg atgctttacc attctctttagttgttaccg taattctgct 3900 ttttcatggg agtttgaatc atggaccata acttttcagttatcagatca actaaagaaa 3960 catttgttgt taagcctaat gtgctgacct atgtgcctgcattttttttt taatctagac 4020 atgtttggag tgagagaaag atggaaaaaa gacatggggtagggacgtaa gtggaaatct 4080 atagccacag cctgaagctt tgaccactgc ggttttcagagccctttctc cacactcatt 4140 tcagagcctc ctatggtttg ggaaggaata acacactggcccattagtaa gggtgaaggc 4200 tggagggatt tgttgacttc ttggaattat cagaggtagggtggtcttta gcacaaagac 4260 ttgcatgcag agatccctgg cagaacaccc agagtgcccgtggctcccac cccagggtct 4320 ggccggtgtg ctggatgcat gcccaagggt gctgggcatcactggtcctt gtgaggatgc 4380 ttttaaagtt ttatatttat gtccccaaac ttggaaacaagaactctact ttagcctaac 4440 cctcatgtcc ttttttgaat tgagaaaatt acaggaaatggtgcctttga aaattagaaa 4500 acttgcttac agagctgttc taaatggtaa atcctcaatttccccaagac cggttgctct 4560 gagagtagct ggtaaagagg ggcgaactaa agacctgtccacctgtagct ccgctcattt 4620 cttagaaacc acctgcttcc cagagtgcca agccacaagtaccaggcttc gtgggcacag 4680 acacctcctg ggctgggcag agtgacagtg ctagaagaccccagagagag ggcaagggct 4740 ttgggcaaga agcactggtg gtgttttagg gaccgtccttctcccctacc cagggaactg 4800 gacctggcag ggcaccgtgc tcatgtggct ccaaggacaagcatggcggt ggccccttct 4860 gccttccagg agaggtcttg cttttgaaac caaaatcatgttcttctaaa gtgtcatctt 4920 ctgccctccc tgtcccaata gggaccacat cttatttgtctcaaacaggg acttgtgagt 4980 acttggcaag ttttgcagcc tatttttgta ttcttaatttggggagtaaa gatgtttggt 5040 ctcaaaaacc tttgaggaat tgccaagaat ggcgagtgattgctttcctt cagagaacag 5100 acacttggaa tttctccttt tagtgtttat atacgtgcagattaatttat atatatatat 5160 atacacacac acatatacag tataaatact catttgattctcgtaaaacg cgcatctggc 5220 gtgtgcagtt gagaaacttg gtggcacatg ggtgttgggggagtagcctg tgttggaggg 5280 acaccagtgc actaggcagc tggggcggcc caggctgaagccatctccgg gtgtctgaga 5340 aaccacccag tgcctcacct ccagatcctg ctggcatcacctccagagcc ctgcatgcac 5400 tggctgaaga gttggtctgt ggagaggatt ttcttggttacttgtattca cggttaattt 5460 acaacccaaa cagcaaaaca cagttggtgg acaagttcatgcaggaccta cagtgaccca 5520 gccatgggca ctagctcatc tttcaggtgg aaaagtacagtgctgcctgc cctggtatgt 5580 ttttcttata gatgttagcc ctgcccaaca gccagggctacactacaaaa ggcaagaatg 5640 cccatgtaag gagcccagca gtctggacag atccttcttcctctgctgtt ggatgagagt 5700 gagtgagtat gctctggacc ttatccttga aagatgatcaaaagcgatga tgagggaggc 5760 agtcatcacg caggtgctta aagggacatt gtaggaggtactcaagggtt tgggggcaaa 5820 accctgaatc cagccagtcg tgcacagaga cacacccacactagcccagt ggcagtgggg 5880 gatgtgggat ggcagcagcc aggtatgtag ccctgtcacaggacagctca ctgtggtttt 5940 gcacactgcc taagggttaa attgtgttgt tgccttcagtagaaggcatt tgtgggctgc 6000 agagttgaga gttgggtgag gttagtctct cctgaagaaaaagcctataa aaagtggcta 6060 atcttatccc ttttctctgt atgcagttgg actcgtcagagatagtaaaa tcatctttta 6120 gtgttttttt gttgctgatg tcttgtaccc atttgttttttacatggggt tgtagatcga 6180 gttctcaaag gtgaaaccag atgatcattc tgataaaggaaatttaaatt tgatacatat 6240 gctttgtata ttttgattac ttgttttcgt ttttgactataaaggagctt ttttattttg 6300 ggaggggagg agtgtcattt ttgagaatct tgggttcctgaaaaagaacg ccctagttgg 6360 atggcttgcc agggccttgg ggtttggtag tgattgtacaacttaaagct cctttctctt 6420 ggctgagtga caggtggctg ttcaggtgga ccaaagcaccttgacacaag gactccacac 6480 tgtgctctct agtagcacaa ggaggaagtt ggacagaacattgggtagtg ccttgcgggc 6540 tcacacatgt actagtggtc tcatctccag ctagccttgggaggccgtcc caccaggaaa 6600 tctctctatt ccgtagcctg agatgtgccc ttgtgggttttatcctgctc agtcagtggc 6660 ctaggggcag gtcctgtgtt ctctccctct ctctcctggctctgggacat ctgtccctgg 6720 ctgcctttca tggaggaagg acactggcct ttctggttggatgctgtgtg gatgctctct 6780 gcttgagcct cgtggtctct tgtctctttt tggaccaatatcctcagatt ggtgcagctt 6840 tttcagttca gatatcacac cccaagtgga taaaggcaacttgcaggaga ggagagccag 6900 ccaagaagaa aattttaaaa ccaaacctcg tttaggattttcctaaagtc atcttctctt 6960 ttttcttgct cagagtttac ctgggagatt tcaccagtttgactcaccat ttgcagatgt 7020 gcttttgtat taaatttaaa ttttcacata tcacatccattctcaaggta gttatatgct 7080 ggagaagaaa aatcctctag acacatgaag gcccacatagtcaagtcttc cagggcaaag 7140 ccagcagccc acccaggtca ggtagccagc agggctcagttcccctcact ccagacacgg 7200 accctcctct tcagggtctc ttgacccagc ttccttctctccttttacct gagagcacag 7260 acctctctca gccagcctgc ccagaccacg gggggctactcccatgtagt ttggggagca 7320 cttgatctca gaaaagctcc attgtctgag caaatgggcagttgtggagc tcaagccttt 7380 ctcctgtgct caagtccctt ccccaagcaa ggcttcaacctcatctaccc accatgtagt 7440 tttctctggc catttaagtg gggcggcagg gacatggttgggccatgcca caccagggct 7500 ggtgaggcaa ccagttttga ttttgacaga gtggctggaggaaaagtggc aatcaaggtg 7560 ctgcttggtt tgctctgagt gcaaatggaa ccaacaggtttctgctgcaa tctgtgtgtt 7620 cccagtgcca ggtcacacca ggaggggtgg ggcagggctaaccaagtggt ctctgaactc 7680 accgagcgtc tgcacttggt tgtgaagtta atgggagtacagagagcgtc tggccttgga 7740 gaggggttga gagcctcctt tttggttctt cattcctgagctcttgcctg cccacaaatc 7800 tgacctcttt gaatggggac gcagtccttc aacagagaagtttctatggc aaagaagttt 7860 ctatttagct ctagatccag cagagtcatc cattctaactgccctgaagt ctagagcagg 7920 ggagggaacc cagaggctgg ggatgagact aggcagaccctggttaccat atggacaagg 7980 acaggggaaa gcaccccctt cctcaatttc tgaaagttctatctttgggt tcgcaggact 8040 ttgaggatga taaagaacat ataggtacta gcttgttgttgctggtccaa agcttccaca 8100 gccctgagaa tttggctttc gtggctgctc tggcagctgagcgaagggag gaaggcagcc 8160 gctctggtgg ggactctagg caccttccct gctgtccacttggataggcg gtgagcccca 8220 gggtactgag aggagcctga gcatttacct gccattagtgcctcttcctt caggagactg 8280 gcttgaaacg tgtgttcatg tgcgcgtgca cacacacacatgagcacctg tatgtgttaa 8340 tgaatagttt ttcttggtta atgcttttta acttctgttcctttccgtaa gtggatgatt 8400 caaaattaac gtgacttggc tgggcgcagt ggctcacacctgtaatccca gcactttggg 8460 aggccaaggc cacccagata aacagccaca ggtcagaaggctgctgagtg cccctggaag 8520 cagaatagct gggcaatggg tccttgactc tctgaaatctcctcatttac tgctgaaagg 8580 ggaaaatgac aagaatagtt tatcccagaa agcatttctctacgttgctc atgttttgga 8640 tgagtctgag agaggcgtgc tggtcaccat gacaacagagacaggccccg actctgaggt 8700 gaagaaagct caggaggagg ccccgcagca gcccgaggctgctgccgctg tgaccacccc 8760 tgtgacccct gcaggccacg gccacccaga ggccaactccaatgagaagc atccatccca 8820 gcaggacacg cggcctgctg aacagagcct agacatggaggagaaggact acagtgaggc 8880 cgatggcctt tcggagagga ccacgcccag caaggcccagaaatcgcccc agaagattgc 8940 caagaaatac aagagtgcca tctgccgggt cactctgcttgatgcctcgg agtatgagtg 9000 tgaggtggag aaacatggcc ggggccaggt gctgtttgacctggtctgtg aacacctcaa 9060 cctcctagag aaggactact tcggcctgac cttctgtgatgctgacagcc agaagaactg 9120 gctggacccc tccaaggaga tcaagaagca gatccggagtgagtggcttg ttgtgtttgg 9180 ggaggtgggt agcccctgga attttgcctt cacagtcaagttctacccgc ctgatcctgc 9240 ccagctgaca gaagacatca caagatacta cctgtgcctgcagctgcggg cagacatcat 9300 cacgggccgg ctgccatgct cctttgtcac gcatgccctactgggctcct acgctgtgca 9360 ggctgagctg ggtgactatg atgctgagga gcatgtgggcaactatgtca gcgagctccg 9420 cttcgcccct aaccagaccc gggagctgga ggagaggatcatggagctgc ataagacata 9480 tagggggatg accccgggag aagcagaaat ccacttcttagagaatgcca agaagctttc 9540 catgtacgga gtagacctgc accatgccaa ggactctgagggcatcgaca tcatgttagg 9600 cgtttgtgcc aatggcctgc tcatctaccg ggaccggctgagaatcaacc gctttgcctg 9660 gcccaagatc ctcaagatct cctacaagag gagtaacttctatatcaaga tccggcctgg 9720 ggagtatgag caatttgaga gcacaattgg ctttaagctcccaaaccacc ggtcagccaa 9780 gagactgtgg aaggtctgca tcgagcatca tacattcttccggctggtgt cccctgagcc 9840 cccacccaag ggcttcctgg tgatgggctc caagttccggtacagtggga ggacccaggc 9900 acagactcgc caggccagcg ccctcattga ccggcctgcacccttctttg agcgttcttc 9960 cagcaaacgg tacaccatgt cccgcagcct tgatggagcagagttctccc gcccagcctc 10020 ggtcagcgag aaccatgatg cagggcctga cggtgacaagcgggatgagg atggcgagtc 10080 tggggggcaa cggtcagagg ctgaggaggg agaggtcaggactccaacca agatcaagga 10140 gctaaagttc ttagacaagc cagaagatgt cttgctgaagcaccaggcca gcatcaatga 10200 gctcaaaagg accctgaagg agcccaacag caaactcatccaccgggatc gagactggga 10260 acgggagcgc aggctgccct cctcccccgc ctccccctcccccaagggca cccctgagaa 10320 agccaatgag tcccagagga cccaggacat ctctcagcgggacttggtac ctgagcctgg 10380 agcagccgca ggcttggaag tgttcactca gaaaagcctcgcagcatctc ctgagggttc 10440 agagcattgg gtatttatag aaagagagta cactaggccagaagagctcg gtctcctaaa 10500 agtgaccacc atgcagcagg aagaaaggca ggcaggccttgctggtatcc ttgccaacgg 10560 cagactctcc aaggtagacg ttctggtgga caagttcaaagtggaagtgg ccacagaaga 10620 aatggtggga aacagaagag caaacaccca gcaacaaggaaaaatgattg caagtcctga 10680 agactttgag tcagtggggg aggaaggccc ctggatcagggaaagcccag gaggggctgc 10740 cctggcttcc ggccgcacat tggcagaaaa gctcctcgagggctctgagc tcagggcaga 10800 caccagagag gcaaccatca ggaaccgctg catgtcagatggtcagccgg agggccagac 10860 agagctgagg aaggggctgg aggagcctca cacttgtgggagacccactg ctccagggac 10920 caggccagca gaggtggacg tcctctctcc agcctccgacaagggaggac tccagtcgtt 10980 tctattggat ccagcccacg cagaagccag agctgagttgagcaatgaaa ctgatacttc 11040 ctttgcagag aggagcttct atttaaatta tgaagaaaaagactcagaag accaagtcct 11100 ccctccaccc ctggaggaga gaaaagggcg cctggatgcccctcccggag gtgagcccag 11160 gccgacgctg aattccttag acctgagggt ttctgctgctgcttccagca ggagcaagga 11220 cgaagcccac atgacttccc caaaggaagg ggcagggacccccaagaacc atggaggacc 11280 tggtgacctg aagggatctc ccgcaggaca gacgtttgctgaaggctggg aagatgccca 11340 gtggggagtg gaaggagagt ttccccacct gacagccagcgcagcccgag aggaagggac 11400 ccccgtgagt ggagatttgc tgggaaaggc tgaggaaagtcccacagagg aactgaagaa 11460 gcaccctcct cacagaggac agggcgtgca tcccgacccccaggcctgcg cccttcctcg 11520 ggccatccct ctgaatgtca ggaagccagt caaaccagacagaggcaact tcccacccaa 11580 agagagggga gtggttccca cccagaaagg aggggctgagctgaaggacc gcgaggcttc 11640 agcatttctt cacatggagg tgatcattcc cctgccagcctcccctggtc attctgagga 11700 cctggcagct ctggaggaag cttctccaag cccaacctcccatgggtcag gggagccttc 11760 ggagctcagg gagccctttc ttagacatgt ccatctttcgaaagccagcc cagagcccaa 11820 ggaccaagta gggtttgtgg tgtcccctgc cacaggaggtgagcgcaggc ctcctcccat 11880 caccagcaga aagcccagag tagtccctga agaagctgaggggcgcatac ctctggggtt 11940 tgggttccct tcagggaagc gaagggagat gacctctttccaggctgggg accaagaggg 12000 ctccctagaa gatattagca agacctcagt ggccaacaaaattcggatat ttgagaccca 12060 cggagctgaa actcgccgaa tgagtgaggg tgaagcaaggtcccttccaa atgacgtatc 12120 ttcagaggca cccgtgggac aagcagagca gcagcggagtacgctctcag acctgggctt 12180 cgcccaactc cagcccccag gggactttgc cagccccaaagccacacatt ccacagtgat 12240 acctctggct accagacact tcagggagga cacttctgcatcctaccagg aagcacacac 12300 ggaactagag cccgtgtccc ccaattcagg ctgtgaaaccacgctggcag aagctactgg 12360 aactggggta actggccgca acaaatccgg agatgcggtcagggaagaga agcgctccac 12420 caacttagca gccaacaccc ctgggaaggg ggggcgcctgagatttgcca gcccctcggg 12480 ccctcagaga gcagggctga gggagggctc cgaggagaaagtcaaaccac cacgtccccg 12540 ggccccagag agtgacacag gcgatgagga ccaggaccaggagagggaca cggtgttcct 12600 gaaggacaac cacctggcca ttgagcgcaa gtgctccagcatcacggtca gctctacgtc 12660 tagcctggag gctgaggtgg acttcacggt cattggtgactaccatggca gcgccttcga 12720 agacttctcc cgcagcctgc ctgagctcga ccgggacaaaagcgactcgg acactgaggg 12780 cctgctgttc tcccgggatc tcaacaaggg ggcccccagccaggatgatg agtctggggg 12840 cattgaggac agcccggatc gaggggcctg ctccaccccggatatgcccc agtttgagcc 12900 cgtgaaaaca gaaaccatga ctgtcagcag tctggccattagaaagaaga ttgagccgga 12960 ggccgtactg cagaccagag tctccgctat ggataacacccaggttgatg ggagtgcctc 13020 agtggggagg gagttcatag caaccactcc ctccatcaccacggagacca tatcgaccac 13080 catggagaac agtctcaagt ccgggaaggg ggcagctgccatgatcccag gcccacagac 13140 ggtggccacg gaaatccgtt ctctttctcc gatcatcgggaaagatgtcc tcaccagcac 13200 ctacggcgcc actgcggaaa ccctctcaac ctccaccaccacccatgtca ccaaaactgt 13260 gaaaggaggg ttttctgaga caaggatcga gaagcgaatcatcattactg gggatgaaga 13320 tgtcgatcaa gaccaggccc tggctttggc catcaaggaggccaaactgc agcatcctga 13380 tatgctggta accaaagctg tcgtatacag agaaacagacccatccccag aggagaggga 13440 caagaagcca cagaagctaa aacgagaaac taagaataacaaaaggcaac cttgcatccg 13500 gcggaacctg caggcacggg ccattcttcc cgcgacccagggctctgccg gaccgcttcc 13560 cccgtcgctc cagtcagacg ccaaagctgg agaacttccggtgcgtttcc gctgtaccgg 13620 aacgtggggc gaggcgctgt tcatcaaaga aaaagggttcttttggtcac ccaccactgg 13680 ccccatggct gccgtgcaga tggatcctga gctagccaagcgcctcttct ttgaaggggc 13740 cactgtggtc atcctgaaca tgcccaaggg aacagagtttgggattgact ataactcctg 13800 ggaggtcggg cccaagttcc ggggcgtgaa gatgatccctccaggcatcc acttcctcca 13860 ctacagctct gtggacaagg ctaatccgaa ggaagtaggccctcgtatgg gtttcttcct 13920 tagcctgcac cagcgggggc tgacagtgct gcgctggagcacactcaggg aagaggtaga 13980 cctgtcccca gccccagagt ctgaggtgga ggccatgagggccaacctcc aggagctgga 14040 ccagttcctg gggccttacc catatgccac cctgaagaagtggatctcac tcaccaactt 14100 catcagcgaa gccacagtgg agaagctaca gcccgagaatcgacagatct gtgccttttc 14160 cgatgtgcta cctgtgctct ccatgaagca caccaaggaccgcgtggggc agaatctacc 14220 ccgctgtggc attgagtgca aaagctacca agagggcctggcccggctac cagagatgaa 14280 gcccagagcc gggacagaga tccgcttctc agagctgcccacgcagatgt tcccagaggg 14340 tgccacgcca gctgagataa ccaagcacag catggacctgagctatgccc tggagactgt 14400 gctcaacaag cagttcccca gcagccccca ggatgtgcttggtgaactcc agtttgcttt 14460 tgtgtgcttc ctgctgggga atgtgtacga ggcatttgagcattggaagc ggctcctgaa 14520 cctcctgtgc cggtcagaag cagccatgat gaagcaccacaccctctaca tcaacctcat 14580 ctccatcctg taccaccagc ttggtgagat ccccgctgacttcttcgtag acattgtctc 14640 ccaagacaac ttcctcacca gcaccttaca gaaaggattcttgattgaca tcagtgaaga 14700 cttcatcgag gtcctgtttc tcagaggtac agaacgcttttctaaagcca acctactgtt 14760 ggatgagtcc ctcattgccg tggatgccac cctgagaaagaaagctgaaa agttccaagc 14820 tcacctgacc aagaagttcc ggtgggactt tgctgcggaacctgaggact gtgccccggt 14880 ggtggtggag ctccctgagg gcatcgagat gggctaa14917 12 1823 DNA Homo sapien 12 cccacttccg gagacctcac acaagatggcggcacccgag gaacacgatt ctccgaccga 60 agcgtcccag cccgattgtg gaagaggaggaaactaaaac atttaaagac ctgggtgtga 120 cagatgtgtt gtgtgaagct tgtgaccagttgggatggac aaaacccacc aagatccaga 180 ttgaagctat tcctttggcc ttacaaggtcgtgatatcat tgggcttgca gaaactggct 240 ctggaaagac aggcgccttt gctttgcccattctaaacgc actgctggag accccgcagc 300 gtttgtttgc cctagttctt accccgactcgggagctggc ctttcagatc tcagagcagt 360 ttgaagccct ggggtcctct attggagtgcagagtgctgt gattgtaggt ggaattgatt 420 caatgtctca atctttggcc cttgcaaaaaaaccacatat aataatagca actcctggtc 480 gactgattga ccacttggaa aatacgaaaggtttcaactt gagagctctc aaatacttgg 540 tcatggatga agccgaccga atactgaatatggattttga gacagaggtt gacaagatcc 600 tcaaagtgat tcctcgagat cggaaaacattcctcttctc tgccaccatg accaagaagg 660 ttcaaaaact tcagcgagca gctctgaagaatcctgtgaa atgtgccgtt tcctctaaat 720 accagacagt tgaaaaatta cagcaatattatatttttat tccctctaaa ttcaaggata 780 cctacctggt ttatattcta aatgaattggctggaaactc ctttatgata ttctgcagca 840 cctgtaataa tacccagaga acagctttgctactgcgaaa tcttggcttc actgccatcc 900 ccctccatgg acaaatgagt cagagtaagcgcctaggatc ccttaataag tttaaggcca 960 aggcccgttc cattcttcta gcaactgacgttgccagccg aggtttggac atacctcatg 1020 tagatgtggt tgtcaacttt gacattcctacccattccaa ggattacatc catcgagtag 1080 gtcgaacagc tagagctggg cgctccggaaaggctattac ttttgtcaca cagtatgatg 1140 tggaactctt ccagcgcata gaacacttaattgggaagaa actaccaggt tttccaacac 1200 aggatgatga ggttatgatg ctgacagaacgcgtcgctga agcccaaagg tttgcccgaa 1260 tggagttaag ggagcatgga gaaaagaagaaacgctcgcg agaggatgct ggagataatg 1320 atgacacaga gggtgctatt ggtgtcaggaacaaggtggc tggaggaaaa atgaagaagc 1380 ggaaaggccg ttaatcactt ttatgaaggctcgagttctg ctgttctgta aaagagaatt 1440 ggagaatgaa acctgctcca acagagatcatgagactgaa attggtcaga attgtgtcca 1500 gaatgtgctc agctaattca gtattcttccccattctggg ttggagttta ctgcagagta 1560 attcttacag tgctgatgtc aagactgttactgttcttcg actttgattc cttgctcatg 1620 acatgagtag ggtgtgctct tctgtcacttcacacagacc ttttgccttt tttagctgca 1680 agtcaaggac taggttgatg atgcccatgacctgtaattg taaagaagct tggacatctg 1740 caaatgatat ttaaaccatc ttggcttgtgctttattcaa actaatgtga aacaataaat 1800 ttaaatatta tttttaaaaa aaa 1823 13869 DNA Homo sapien 13 cagcattgca gcagctccac catggcctgg gctcctctgctcctcaccct cctcagtctc 60 ctcacagggt ccctctccca gcctatcttg actcagccaccttctgcatc agcctccctg 120 ggagcctcgg tcacactcac gtgcagtgtg agcagcgactacaagaatct tgaagtggac 180 tggtttcagc agagaccagg gaagggcccc cgttttgtcatgcgagtggg cactggtggc 240 gttgtgggat tcagaggggc tgacatccct gatcgcttttcagtctcggg ctcaggcctg 300 aatcggtttc tgaccatcag gaacatcgaa gaagaggatgagagtgacta ccactgtggg 360 acggaccttg gcagtgggac cagcttcgtg tcttgggtgttcggcggagg gaccaagttg 420 accgtcctaa gtcagcccaa ggctgccccc tcggtcactctgttcccgcc ctcctctgag 480 gagcttcaag ccaacaaggc cacactggtg tgtctcataagtgacttcta cccgggagcc 540 gtgacagtgg cctggaaggc agatagcagc cccgtcaaggcgggagtgga gaccaccaca 600 ccctccaaac aaagcaacaa caagtacgcg gccagcagctatctgagcct gacgcctgag 660 cagtggaagt ccaacagaag ctacagctgc caggtcacgcatgaagggag caccgtggag 720 aagacagtgg cccctacaga atgttcatag gttctaaaccctcacccccc ctacgggaga 780 ctagagctgc aggatcccag gggaggggtg tctcctcccacccgcaaggc gtcaagccct 840 tctccctgca ctcaataacc gatcgaata 869 14 799 DNAHomo sapien 14 ccctgctcag ctcttggggc cgctaatgct ctgggtccct gtgcagagattgtgatgacc 60 cagactccac tctccttgtc tatcacccct ggagagcagg cctccatgtcctgcaggtct 120 agtcagagcc tcctgcatag tgatggatac acctatttgt attggtttctgcagaaagcc 180 aggccagtct ccacagctcc tgatctatga agtttccaac cggttctctggagtgtcacc 240 attaggttca gtggcagcgg gtccgggaga gaattcacat tgagaatcagccgggtggag 300 gctgacgatg ctggagttta ctactgcatg caaactacac agactccgaacacttttggc 360 caggggacga ggctggagat caaacgaact gtggctgcac catctgtcttcatcttcccg 420 ccatctgatg agcagttgaa atctggaact gcctctgttg tgtgcctgctgaataacttc 480 tatcccagag aggccaaagt acagtggaag gtggataacg ccctccaatcgggtaactcc 540 caggagagtg tcacagagca ggacagcaag gacagcacct acagcctcagcagcaccctg 600 acgctgagca aagcagacta cgagaaacac aaactctacg cctgcgaagtcacccatcag 660 ggcctgagct cgcccgtcac aaagagcttc aacaggggag agtgttagagggagaagtgc 720 ccccacctgc tcctcagttc cagcctgacc ccctcccatc ctttggcctctgaccctttt 780 tccacagggg acctacccc 799 15 1731 DNA Homo sapien 15tttttttttt ttggttgtca ttgaggatat ttattggggt ttcatgagtg cagggagaag 60ggctggatga cttgggatgg ggagagagac ccctcccctg ggatcctgca gctccaggct 120cccgtgggtg gggttagagt tgggaaccta tgaacattct gtaggggcca ctgtcttctc 180cacggtgctc ccttcatgcg tgacctggca gctgtagctt ctgtgggact tccactgctc 240gggcgtcagg ctcaggtagc tgctggccgc gtacttgttg ttgctctgtt tggagggttt 300ggtggtctcc actcccgcct tgacggggct gccatctgcc ttccaggcca ctgtcacagc 360tcccgggtag aagtcactga tcagacacac tagtgtggcc ttgttggctt ggagctcctc 420agaggagggc gggaacagag tgacagtggg gttggccttg ggctgacctg tgtggacagg 480gaagggggtg agagagggca gacagaatac cggggtgttg tggagcccct ctctctgtct 540aaagtctctg ggagggttca cagtgtggcc atccggtcca cccggggttc tctcctcttc 600tttccccatc ctttccactc atgccctgtg gagagcagac agctctgtgc cttcctagga 660gccctcccaa gtcacctttc acaggtgtcc tggcccagcc cctctcctct acagcctcaa 720tttctccata tacccagggc aggctgtgtt ccttcttctc tgtgttcctt cttctctgat 780ctctggagtc tgagtttaga atctggcctt gacccctgga tccctcattt ccatccctat 840acccccctcc atcacccact ttattcttcc aggacctaga gcctcctccg tgaactgtgg 900gtcccctgct catcctgtag gactgtcctg gcaggcagtg tgtggggaga cccaagcctg 960ctttgaactg gagcttccta tccctcacag gcaccgggct ctgccccagc ccagcccact 1020tggctctcct ggcaaggagt gggctccacc tagacaagcc tcagggctcc tctgaggctc 1080tgagatgttt cctgtctcca caagcccgac cacagaaccc ttcacctcag ctgttcctgg 1140ggctgctccc acagactatg agactcagca gcccccaggc ccaccccagc agcctgtgat 1200gaccccaaac ttcacctggt agccatggag ttctctgcac ccctcattca ctcccctctg 1260gtcatttcct gagtctaaca tgccctttga ggaaggcagg aggccgatcc gtgaacagag 1320agacactggg ccccagaggt gacggggctc cagggacaga cacatctctg ccctaagaga 1380ctgtctcctt tctggtgact gtcctgggag ggttggattc tggcacctca cccagtctca 1440cccgtccctc tgtgtcccca tgtcctcatg acccagcaca ggccacagag ctgcagccta 1500gacccagagc cctctctgtg ccctccattg gtctcccctt ggggtgacct ctgtgtcacc 1560aggccgtgtg gccctcccag gctgattggc atccagtcct cagcctagac cctcagctgt 1620tcttggggct gctcccctag actatgagac tcagcagctc ccaggcccac cccagcagcc 1680tctgtccttg accccaggag tcactgggca atgtccctaa ggacactgca g 1731 16 662 DNAHomo sapien 16 gccaaagagt tccaggccca caagaggact ggctatgagg aagagacctggaatctgaag 60 gaatgtgttg ggcgttgtgc aaaccctaac gtaaatttcc tgacaaaggtagaaagccct 120 ggcatggttc agaggtgggg cctcctccta tgtcgacggg attctagattcacaccatgg 180 cagaaaatct atttcaggaa cttcaggaac attttcaagc tctgacggcaacattaaacc 240 tcagaaatat ccttttctac ctttaacaaa tgctgtgatt ctttcggactggtagattat 300 catggagtat ctttttgttg tctggtagta gtaggtaata gtttacttaggatttcccag 360 tatttacttc tgtgctttta tgtggcttcc tgatgtgtta attacccctcacctatagca 420 aaagctgtac ctccggccgg gtgcagtggc ccataaggaa attttctaactatatgaact 480 gaagggccat ggcttctaca gaaaatatgt ttacatcaaa tacaatcttgggaagaataa 540 aaaatagctc tcctattcct tacagggaag gctataaaca atattttattgtactgtttt 600 tataaccaga gtaaaccttt ggattctgtc atggattgaa ttgtgtaaccgcaaaattta 660 ta 662 17 336 DNA Homo sapien misc_feature (268)..(268)a, c, g or t 17 tttaaaaaaa atccatctta gtatcttgac ccccaccctt cacccactcacagagaagcc 60 cacatgagga aacaggttat gtcttggaca tctctgtccc cctcagtgtctggtatagtg 120 actgacacac agcatgttct caagaaatgt ttgaatcaca gtacattgaatcagtaacag 180 tctgactgac ccccaggcag aaaatgcaga ggcatttttt ctctctattccagatttcag 240 ctgtagcctc ttgtaattct catattgntt ttcaatcacc agaattgatttccctcatcc 300 ctcttcccag ggtcatctcc agtgaactgt attaat 336 18 3300 DNAHomo sapien misc_feature (892)..(892) a, c, g or t 18 gagcccgagccgcgccaccc cgcctggcca tggcttttgc aagtttccgc cgcatcctgg 60 ccttgtctaccttcgagaag agaaagtccc gcgaatatga gcacgtccgc cgcgacctgg 120 accccaacgaggtgtgggag tatcgtgggc gagctgggcg acggcgtctt tcggaatggt 180 ttacaaggccaagaataagg agacgggtgc tttggctgcg gcaatagtca ttgaaaccaa 240 gagtgaggaggagctggagg actacatcgt ggagattgag atcctggcca cctgcgacca 300 cccctacattgtgaagctcc tgggagccta ctatcacgac gggaagctgt ggatcatgat 360 tgagttctgtccagggggag ccgtggacgc catcatgctg gagctggaca gaggcctcac 420 ggagccccagatacaggtgg tttgccgcca gatgctagaa gccctcaact tcctgcacag 480 caagaggatcatccaccgag atctgaaagc tggcaacgtg ctgatgaccc tcgagggaga 540 catcaggctggctgactttg gtgtgtctgc caagaatctg aagactctac agaaacgaga 600 ttccttcatcggcacgcctt actggatggc ccccgaggtg gtcatgtgtg agaccatgaa 660 agacacgccctacgactaca aagccgacat ctggtccctg ggcatcacgc tgattgagat 720 ggcccagatcgagccgccac accacgagct caaccccatg cgggtcctgc taaagatcgc 780 caagtcagaccctcccacgc tgctcacgcc ctccaagtgg tctgtagagt tccgtgactt 840 cctgaagatagccctggata agaacccaga aacccgaccc agtgccgcgc antgctggag 900 catcccttcgtcagcagcat caccagtaac aaggctctgc gggagctggt ggctgaggcc 960 aaggccgaggtgatggaaga gatcgaagac ggccgggatg agggggaaga ggaggacgcc 1020 gtggatgctgcctccaccct ggagaaccat actcagaact cctctgaggt gagtccgcca 1080 agcctcaatgctgacaagcc tctcgaggag tcaccttcca ccccgctggc acccagccag 1140 tctcaggacagtgtgaatga gccctgcagc cagccctctg gggacagatc cctccaaacc 1200 accagtcccccagtcgtggc ccctggaaat gagaacggcc tggcagtgcc tgtgcccctg 1260 cggaagtcccgacccgtgtc aatggatgcc agaattcagg tagcccagga gaagcaagtt 1320 gctgagcagggtggggacct cagcccagca gccaacagat ctcaaaaggc cagccagagc 1380 cggcccaacagcagcgccct ggagaccttg ggtggggaga agctggccaa tggcagcctg 1440 gagccacctgcccaggcagc tccagggcct tccaagaggg actcggactg cagcagcctc 1500 tgcacctctgagagcatgga ctatggtacc aatctctcca ctgacctgtc gctgaacaaa 1560 gagatgggctctctgtccat caaggacccg aaactgtaca aaaaaacctc aagcggacac 1620 gcaaatttgtggtggatggt gtggaggtga gcatcaccac ctccaagatc atcagcgaag 1680 atgagaagaaggatgaggag atgagatttc tcaggcgcca ggaactccga gagcttcggc 1740 tgctccagaaagaagagcat cggaaccaga cccagctgag taacaagcat gagctgcagc 1800 tggagcaaatgcataaacgt tttgaacagg aaatcaacgc caagaagaag ttctttgaca 1860 cggaattagagaacctggag cgtcagcaaa agcagcaagt ggagaagatg gagcaagacc 1920 atgccgtgcgccgccgggag gaggccaggc ggatccgcct ggagcaggat cgggactaca 1980 ccaggttccaagagcagctc aaactgatga agaaagaggt gaagaacgag gtggagaagc 2040 tcccccgacagcagcggaag gaaagcatga agcagaagat ggaggagcac acgcagaaaa 2100 agcagcttcttgaccgggac tttgtagcca agcagaagga ggacctggag ctggccatga 2160 agaggctcaccaccgacaac aggcgggaga tctgtgacaa ggagcgcgag tgcctcatga 2220 agaagcaggagctccttcga gaccgggaag cagccctgtg ggagatggaa gagcaccagc 2280 tgcaggagaggcaccagctg gtgaagcagc agctcaaaga ccagtacttc ctccagcggc 2340 acgagctgctgcgcaagcat gagaaggagc gggagcagat gcagcgctac aaccagcgca 2400 tgatagagcagctgaaggtg cggcagcaac aggaaaaggc gcggctgccc aagatccaga 2460 ggagtgagggcaagacgcgc atggccatgt acaagaagag ccccctgtaa actggagttg 2520 ctggtgggcagccctgcttc ctgcatggaa ctggagctgt atggagttga cgacaagttc 2580 tacagcaagctggatcaaga ggatgcgctc ctgggctcct accctgtaga tgacggctgc 2640 cgcatccacgtcattgacca cagtggcgcc cgccttggtg agtatgagga cgtgtccccg 2700 gtggagaagtacacgatctc acaagaagcc tacgaccaga ggcaagacac ggtccgctct 2760 ttcctgaagcgcagcaagct cggccggtac aacgaggagg agcgggctca gcaggaggcc 2820 gaggccgcccagcgcctggc cgaggagaag gcccaggcca gctccatccc cgtgggcagc 2880 cgctgtgaggtgcgggcggc gggacaatcc cctcgccggg gcaccgtcat gtatgtaggt 2940 ctcacagatttcaagcctgg ctactggatt ggtgtccgct atgatgagcc actggggaaa 3000 aatgatggcagtgtgaatgg gaaacgctac ttcgaatgcc aggccaagta tggcgccttt 3060 gtcaagccagcagtcgtgac ggtgggggac ttcccggagg aggactacgg gttggacgag 3120 atatgacacctaaggaattc ccctgcttca gctcctagct cagccactga ctgcccctcc 3180 tgtgtgtgcccatggccctt ttctcctgac cccattttaa ttttattcat tttttccttt 3240 gccattgatttttgagactc atgcattaaa ttcactagaa acccagaaaa aaaaaaaaaa 3300 19 349 DNAHomo sapien misc_feature (6)..(6) a, c, g or t 19 ttaaanattt aaanatattaaggtntcntn tncngctcat cttcacagga aaanaattan 60 attccnnaca aacacctttcaantnttacc cnggnaagct gtnngaactg gaccttgaag 120 aaggaaatac tttctgactcattgnaattt gntatagtca nagtaagtaa ggcacaaant 180 ttgaagataa acttggnacagtatgtagtg ccttttgtta tnaaatgcaa ctttgggnga 240 ttactngana ttttttcntcataganttac tannanaagt tttnanaatn ttactgatna 300 ccactgtttt tgattttgctattntttnca aattnactat atatgangt 349 20 4665 DNA Homo sapien 20agcggggagg gccccgagcg gcgcagatag ggaggttggg gctgtgcccc gcggcgcggc 60gcctgccact gcgcaggcgc ctcaggaaga gctcggcatc gcccctcttc ctccaggtcc 120cccttccccg caacttccca cgagtgccag gtgccgcgag cgccgagttc cgcgcattgg 180aaagaagcga ccgcggcggc tggaaccctg attgctgtcc ttcaacgtgt tcattatgaa 240gttattagta atacttttgt tttctggact tataactggt tttagaagtg actcttcctc 300tagtttgcca cctaagttac tactagtatc ctttgatggc ttcagagctg attatctgaa 360gaactatgaa tttcctcatc tccagaattt tatcaaagaa ggtgttttgg tagagcatgt 420taaaaatgtt tttatcacaa aaacatttcc aaaccactac agtattgtga caggcttgta 480tgaagaaagc catggcattg tggctaattc catgtatgat gcagtcacaa agaaacactt 540ttctgactct aatgacaagg atcctttttg gtggaatgag gcagtaccta tttgggtgac 600caatcagctt caggaaaaca gatcaagtgc tgctgctatg tggcctggta ctgatgtacc 660cattcacgat accatctctt cctattttat gaattacaac tcctcagtgt catttgagga 720aagactaaat aatattacta tgtggctaaa caattcgaac ccaccagtca cctttgcaac 780actatattgg gaagaaccag atgcaagtgg ccacaaatac ggacctgaag ataaagaaaa 840catgagcaga gtgttgaaaa aaatagatga tcttatcggt gacttagtcc aaagactcaa 900gatgttaggg ctatgggaaa atcttaatgt gatcattaca agtgatcatg ggatgaccca 960gtgttctcag gacagactga taaacctgga ttcctgcatc gatcattcat actacactct 1020tatagatttg agcccagttg ctgcaatact tcccaaaata aatagaacag aggtttataa 1080caaactgaaa aactgtagcc ctcatatgaa tgtttatctc aaagaagaca ttcctaacag 1140attttattac caacataatg atcgaattca gcccattatt ttggttgccg atgaaggctg 1200gacaattgtg ctaaatgaat catcacaaaa attaggtgac catggttatg ataattcttt 1260gcctagtatg catccatttc tagctgccca cggacctgca tttcacaaag gctacaagca 1320tagcacaatt aacattgtgg atatttatcc aatgatgtgc cacatcctgg gattaaaacc 1380acatcccaat aatgggacct ttggtcatac taagtgcttg ttagttgacc agtggtgcat 1440taatctccca gaagccatcg cgattgttat cggttcactc ttggtgttaa ccatgctaac 1500atgcctcata ataatcatgc agaatagact ttctgtacct cgtccatttt ctcgacttca 1560gctacaagaa gatgatgatg atcctttaat tgggtgacat gtgctagggc ttatacaaag 1620tgtctttgat taatcacaaa actaagaata catccaaaga atagtgttgt aactatgaaa 1680aagaatactt tgaaagacaa agaacttaga ctaagcatgt taaaattatt actttgtttt 1740ccttgtgttt tgtttcggtg catttgctaa taagataacg ctgaccatag taaaattgtt 1800agtaaatcat taggtaacat cttgtggtag gaaatcatta ggtaacatca atcctaacta 1860gaaatactaa aaatggcttt tgagaaaaat acttcctctg cttgtatttt gcgatgaaga 1920tgtgatacat ctttaaatga aaatatacca aaatttagta ggcatgtttt tctaataaat 1980ttatatattt gtaaagaaaa caacagaaat ctttatgcaa tttgtgaatt ttgtatatta 2040gggaggaaaa gcttcctata tttttatatt tacctttaat tagtttgtat ctcaagtacc 2100ctcttgaggt aggaaatgct ctgtgatggt aaataaaatt ggagcagaca gaaaagatat 2160agcaaatgaa gaaatatttt aaggaaacct atttgaaaaa aaaagcaaag accatttgat 2220aaaagcctga gttgtcacca ttatgtctta agctgttagt cttaaagatt attgttaaaa 2280aattcagaag aaaagagaga caagtgctct tctctctatc tatgcttaat gcctttatgt 2340aagttactta gttgtttgcg tgtgcctgtg caagtgtgtt tgtgtgtggt tgtgtggaca 2400ttatgtgatt tactatataa ggaggtcaga gatggactgt ggccaggctt ccacattcct 2460gaagcacaca gatctcagga aaggttattt ttgcacttca tatttgttta ctttctccta 2520actcacaagt taaaatcata acttaatttc attaactttt atcatttaac tctctcatgt 2580ttgttgtaac ccgaggtatc caaatgctgc agaaaaattt atgacccaaa tacaaatctc 2640aatatgactg ggacagaatg aggaatggag atttttgtat ttatctttgg gactttatgc 2700cttacttttt aggctataga atagttaaga aattttaaac aaaatttagt atcttttggt 2760ctttcacacc attcatatgt taagtggcag aatagcctta gtgctacctc cacttttttc 2820tccagtattt gcatcacaga aataatccct ctgtttaaca tgtttgttca gagccaaggg 2880tttattgtga agaactgtca tcctgccttt gctagctggt accttctagt aatcaaaatt 2940aatatgaaga aactaggttg tgacagacta gattatattt agtaggggaa aaattgggct 3000caagaaccat tcatcagtac gtgagacaag cagttaatag tatgatcttt aaagttttga 3060caatataaaa taaacttggt aactgtttta caaatataaa agtataataa atatgcagcc 3120cagttaaata ttgattatct gtgatggtaa agaacaacag tggtgccagt catcaaacat 3180acagtgcgtc ctattgagtc actgctaatt tcttgagcct ggtatttgct gcctattgta 3240tttgtggttg ttgagaggca ttttcaaacc ctgtataaat aatccatgct gttggtcata 3300agttaactgt attaagaaca gtaaaataaa taaaaaccaa tagtactaat tttgctttaa 3360aaaaatttct aatttttttc acataaaaca attatcctaa aggttaatag ttgatcgaaa 3420cagaataata gaaaaattct tctttaattt ccattaaaaa gcaaatagca ttgacacatt 3480taaagctttt catttaaagt agtggatgtt tttgaagtat ctaaaatagt agcagaatat 3540tttatacttg gtccttgcaa tggtgtgagt tttaatgatt gcattatcgt gattggtggt 3600tatgagtttc agaaatctat acttggcatc caactcatga gtggatttta tataggatgg 3660aacaggaagg tatgtcctgt cagtatctta accctttcaa caagacattt acctatttgt 3720ctttccttac gttctcaaaa tattaactcg aattgtaaat taagcaaaaa tgtaaaaagt 3780atatgttgat gggacaagaa gaatagtatt tatttaataa aacatatatt atattgaact 3840atgtgttaat tcatttgtat cttttaaaaa attatcactg ttaaagccat tgactccttt 3900agtacactga gaaaaatctt atagtaaaac tagcctttca cattaaggtt ttggtgtgta 3960ttttgttaaa taactaacat gctgctctat tttctgggtg tagaaagtat ttggctctag 4020gaaacattta cttgtttgtg aaaacaatac cccaaggtaa taggaaaagt ttgagttaag 4080tgtttttaat tcagtcagtg aattcagaat aagtacattc atgtataaca tagggacagt 4140tctgctgctg ttatttatat gcaattcttc tggtaaatag caatagaata aaacatattt 4200caatgtttgt gtataggttt tatattatta ttccactagg aatggcataa gaatttatag 4260ataaattctt gtaacattaa aggattaaaa tgtttttaca ttgtttttgg gtgtctcctt 4320cttgtgccca tatctgataa gctttatgga ttattgcatt taattccttt tatttggagg 4380gttttacttc cttgttaaca tataaagtta taaatgaagg acaaggagga gatggaaaat 4440gtgtatttat tgttaattct taaaatagtg tgtaaataaa ataacatcag tgtgctttaa 4500agaaatgtgt atgtagtgcc ttaatttaaa ttaaaatatt tttgactgtt acttgagttc 4560agaattaatg actttgttca tgatttttaa aatgtgtgtg aataaaatct accaaaaaat 4620tcttactgta attattaaat ataaagttca gtgtcaaaaa aaaaa 4665 21 437 DNA Homosapien 21 tagcaacagg cctggaggtg ctgcagtagt gggggaaaat ggaaggtggagggtggagtg 60 tttgctgcag gacagctgag tggagggtgg ggacaggtgc aaactggagaggcctagaga 120 gctagagaag caagtaaggg ccagggccag agtcggcttc aatggaacaacagcccagtg 180 ccctaaggcc cctaactctt gctggctgtt tcttgacccc aagccagggttgggagtcct 240 ctgggcatcc attttttcta aaggaactgg acagagtaca cacaggaaaggaagctgtca 300 ccctcttgcc atctggctcc aggggcctcc agtccagcat tcctccttcttcccttgatt 360 gggtggggcc acatgatggg cagccaggct ctgggctgtc ccactagagcagctgcaaac 420 acagccatgt ttcagtg 437 22 355 DNA Homo sapienmisc_feature (17)..(17) a, c, g or t 22 caagctggct tctgttnaga tgagctncnnggagatgcta ctgcatggca caggaagaac 60 gtgtgaccac taggattatt tccagcataantggctttgc atggntgaag ttntagcaat 120 gaatttctat aagccctttt aaaattggaattcataaaca agtctctgtg ctctcaccnt 180 gtggcanttn tttctgctct ttttgttgtttnattgtgtt ctcactgcta cctagctagc 240 atcntggtgt catggaagtg gaccagatattttcacaccc attatattct agatgctgtg 300 ttaagatnca ngacaacnan ntngnnncnngatgtaaaat tttntagncn gnagg 355 23 21 DNA Artificial Sequence Synthetic23 ccagagccca aatcttgtga c 21 24 20 DNA Artificial Sequence Synthetic 24gcggctttgt cttggcatta 20 25 21 DNA Artificial Sequence Synthetic 25attgccatcc cagtgacagt g 21 26 21 DNA Artificial Sequence Synthetic 26ttgggagatg tgggtgatga g 21 27 22 DNA Artificial Sequence Synthetic 27cctgccctgg tatgtttttc tt 22 28 21 DNA Artificial Sequence Synthetic 28cagcccacaa atgccttcta c 21 29 24 DNA Artificial Sequence Synthetic 29ccactaggat tatttccagc ataa 24 30 24 DNA Artificial Sequence Synthetic 30ggtgtgaaaa tatctggtcc actt 24 31 18 DNA Artificial Sequence Synthetic 31agccattgcc atcccagt 18 32 20 DNA Artificial Sequence Synthetic 32atgttcttca cgctcttcgc 20 33 24 DNA Artificial Sequence Synthetic 33aggaagtgct ggaagaggct ggct 24 34 19 DNA Artificial Sequence Synthetic 34aagggagcac cgtggagaa 19 35 19 DNA Artificial Sequence Synthetic 35agggctggat gacttggga 19 36 24 DNA Artificial Sequence Synthetic 36ttcccaactc taaccccacc cacg 24 37 7444 DNA Homo sapien 37 gtctcctctggatcttaact actgagcgca atgctgagcc atggagccgg gttggccttg 60 tggatcacactgagcctgct gcagactgga ctggcggagc cagagagatg taacttcacc 120 ctggcggagtccaaggcctc cagccattct gtgtctatcc agtggagaat tttgggctca 180 ccctgtaactttagcctcat ctatagcagt gacaccctgg gggccgcgtt gtgccctacc 240 tttcggatagacaacaccac atacggatgt aaccttcaag atttacaagc aggaaccatc 300 tataacttcaggattatttc tctggatgaa gagagaacag tggtcttgca aacagatcct 360 ttacctcctgctaggtttgg agtcagtaaa gagaagacga cttcaaccag cttgcatgtt 420 tggtggactccttcttccgg aaaagtcacc tcatatgagg tgcaattatt tgatgaaaat 480 aaccaaaagatacagggggt tcaaattcaa gaaagtactt catggaatga atacactttt 540 ttcaatctcactgctggtag taaatacaat attgccatca cagctgtttc tggaggaaaa 600 cgttctttttcagtttatac caatggatca acagtgccat ctccagtgaa agatattggt 660 atttccacaaaagccaattc tctcctgatt tcctggtccc atggttctgg gaatgtggaa 720 cgataccggctgatgctaat ggataaaggg atcctagttc atggcggtgt tgtggacaaa 780 catgctacttcctatgcttt tcacgggctg tcccctggct acctctacaa cctcactgtt 840 atgactgaggctgcagggct gcaaaactac aggtggaaac tagtcaggac agcccccatg 900 gaagtctcaaatctgaaggt gacaaatgat ggcagtttga cctctctaaa agtcaaatgg 960 caaagacctcctggaaatgt ggattcttac aatatcaccc tgtctcacaa agggaccatc 1020 aaggaatccagagtattagc accttggatt actgaaactc actttaaaga gttagtcccc 1080 ggtcgactttatcaagttac tgtcagctgt gtctctggtg aactgtctgc tcagaagatg 1140 gcagtgggcagaacatttcc agacaaagtt gcaaacctgg aggcaaacaa taatggcagg 1200 atgaggtctcttgtagtgag ctggtcgccc cctgctggag actgggagca gtatcggatc 1260 ctactcttcaatgattctgt ggtgctgctc aacatcactg tgggaaagga agaaacacag 1320 tatgtcatggatgacacggg gctcgtaccg ggaagacagt atgaggtgga agtcattgtt 1380 gagagtggaaatttgaagaa ttctgagcgt tgccaaggca ggacagtccc cctggctgtc 1440 ctccagcttcgtgtcaaaca tgccaatgaa acctcactga gtatcatgtg gcagacccct 1500 gtagcagaatgggagaaata catcatttcc ctagctgaca gagacctctt actgatccac 1560 aagtcactctccaaagatgc caaagaattc acttttactg acctggtgcc tggacgaaaa 1620 tacatggctacagtcaccag tattagtgga gacttaaaaa attcctcttc agtaaaagga 1680 agaacagtgcctgcccaagt gactgacttg catgtggcca accaaggaat gaccagtagt 1740 ctgtttactaactggaccca ggcacaagga gacgtagaat tttaccaagt cttactgatc 1800 catgaaaatgtggtcattaa aaatgaaagc atctccagtg agaccagcag atacagcttc 1860 cactctctcaagtccggcag cctgtactcc gtggtggtaa caacagtgag tggagggatc 1920 tcttcccgacaagtggttgt ggagggaaga acagtccctt ccagtgtgag tggagtaacg 1980 gtgaacaattccggtcgtaa tgactacctc agcgtttcct ggctcgtggc gcccggagat 2040 gtggataactatgaggtaac attgtctcat gacggcaagg tggttcagtc ccttgtcatt 2100 gccaagtctgtcagagaatg ttccttcagc tccctcaccc caggccgcct ctacaccgtg 2160 accataactacaaggagtgg caagtatgaa aatcactcct tcagccaaga gcggacagtg 2220 cctgacaaagtccagggagt cagtgttagc aactcagcca ggagtgacta tttaagggta 2280 tcctgggtgtatgccactgg agactttgat cactatgaag tcaccattaa aaacaaaaac 2340 aacttcattcaaactaaaag cattcccaag tcagaaaacg aatgtgtatt tgttcagcta 2400 gtccctggacggttgtacag tgtcactgtt actacaaaaa gtggacaata tgaagccaat 2460 gaacaagggaatgggagaac aattccagag cctgttaagg atctaacatt gcgcaacagg 2520 agcactgaggacttgcatgt gacttggtca ggagctaatg gggatgtcga ccaatatgag 2580 atccagctgctcttcaatga catgaaagta tttcctcctt ttcaccttgt aaataccgca 2640 accgagtatcgatttacttc cctaacacca ggccgccaat acaaaattct tgtcttgacg 2700 attagcggggatgtacagca gtcagccttc attgagggct tcacagttcc tagtgctgtc 2760 aaaaatattcacatttctcc caatggagca acagatagcc tgacggtgaa ctggactcct 2820 ggtgggggagacgttgattc ctacacggtg tcggcattca ggcacagtca aaaggttgac 2880 tctcagactattcccaagca cgtctttgag cacacgttcc acagactgga ggccggggag 2940 cagtaccagatcatgattgc ctcagtcagc gggtccctga agaatcagat aaatgtggtt 3000 gggcggacagttccagcatc tgtccaagga gtaattgcag acaatgcata cagcagttat 3060 tccttaatagtaagttggca aaaagctgct ggtgtggcag aaagatatga tatcctgctt 3120 ctaactgaaaatggaatcct tctgcgcaac acatcagagc cagccaccac taagcaacac 3180 aaatttgaagatctaacacc aggcaagaaa tacaagatac agatcctaac tgtcagtgga 3240 ggcctctttagcaaggaagc ccagactgaa ggccgaacag tcccagcagc tgtcaccgac 3300 ctgaggatcacagagaactc caccaggcac ctgtccttcc gctggaccgc ctcagagggg 3360 gagctcagctggtacaacat ctttttgtac aacccagatg ggaatctcca ggagagagct 3420 caagttgacccactagtcca gagcttctct ttccagaact tgctacaagg cagaatgtac 3480 aagatggtgattgtaactca cagtggggag ctgtctaatg agtctttcat atttggtaga 3540 acagtcccagcctctgtgag tcatctcagg gggtccaatc ggaacacgac agacagcctt 3600 tggttcaactggagtccagc ctctggggac tttgactttt atgagctgat tctctataat 3660 cccaatggcacaaagaagga aaactggaaa gacaaggacc tgacggagtg gcggtttcaa 3720 ggccttgttcctggaaggaa gtacgtgctg tgggtggtaa ctcacagtgg ggatctcagc 3780 aataaagtcacagcggagag cagaacagct ccaagtcctc ccagtcttat gtcatttgct 3840 gacattgcaaacacatcctt ggccatcacg tggaaagggc ccccagactg gacagactac 3900 aacgactttgagctgcagtg gttgcccaga gatgcactta ctgtcttcaa cccctacaac 3960 aacagaaaatcagaaggacg cattgtgtat ggtcttcgtc cagggagatc ctatcaattc 4020 aacgtcaagactgtcagtgg tgattcctgg aaaacttaca gcaaaccaat ttttggatct 4080 gtgaggacaaagcctgacaa gatacaaaac ctgcattgcc ggcctcagaa ctccacggcc 4140 attgcctgttcttggatccc tcctgattct gactttgatg gttatagtat tgaatgccgg 4200 aaaatggacacccaagaagt tgagttttcc agaaagctgg agaaagaaaa atctctgctc 4260 aacatcatgatgctagtgcc ccataagagg tacctggtgt ccatcaaagt gcagtcggcc 4320 ggcatgaccagcgaggtggt tgaagacagc actatcacaa tgatagaccg cccccctcct 4380 ccacccccacacattcgtgt gaatgaaaag gatgtgctaa ttagcaagtc ttccatcaac 4440 tttactgtcaactgcagctg gttcagcgac accaatggag ctgtgaaata cttcacagtg 4500 gtggtgagagaggctgatgg cagtgatgag ctgaagccag agcagcagca ccctctccct 4560 tcctacctggagtacaggca caatgcctcc attcgggtgt atcagactaa ttattttgcc 4620 agcaaatgtgccgaaaatcc taacagcaac tccaagagtt ttaacattaa gcttggagca 4680 gagatggagagcctaggtgg aaaatgcgat cccactcagc aaaaattctg tgatggacca 4740 ctgaagccacacactgccta cagaatcagc attcgagctt ttacacagct ctttgatgag 4800 gacctgaaggaattcacaaa gccactctat tcagacacat ttttttcttt acccatcact 4860 actgaatcagagcccttgtt tggagctatt gaaggtgtga gtgctggtct gtttttaatt 4920 ggcatgctagtggctgttgt tgccttattg atctgcagac agaaagtgag ccatggtcga 4980 gaaagaccctctgcccgtct gagcattcgt agggatcgac cattatctgt ccacttaaac 5040 ctgggccagaaaggtaaccg gaaaacttct tgtccaataa aaataaatca gtttgaaggg 5100 catttcatgaagctacaggc tgactccaac taccttctat ccaaggaata cgaggagtta 5160 aaagacgtgggccgaaacca gtcatgtgac attgcactct tgccggagaa tagagggaaa 5220 aatcgatacaacaatatatt gccctatgat gccacgcgag tgaagctctc caatgtagat 5280 gatgatccttgctctgacta catcaatgcc agctacatcc ctggcaacaa cttcagaaga 5340 gaatacattgtcactcaggg accgcttcct ggcaccaagg atgacttctg gaaaatggtg 5400 tgggaacaaaacgttcacaa catcgtcatg gtgacccagt gtgttgagaa gggccgagta 5460 aagtgtgaccattactggcc agcggaccag gattccctct actatgggga cctcatcctg 5520 cagatgctctcagagtccgt cctgcctgag tggaccatcc gggagtttaa gatatgcggt 5580 gaggaacagcttgatgcaca cagactcatc cgccactttc actatacggt gtggccagac 5640 catggagtcccagaaaccac ccagtctctg atccagtttg tgagaactgt cagggactac 5700 atcaacagaagcccgggtgc tgggcccact gtggtgcact gcagtgctgg tgtgggtagg 5760 actggaacctttattgcatt ggaccgaatc ctccagcagt tagactccaa agactctgtg 5820 gacatttatggagcagtgca cgacctaaga cttcacaggg ttcacatggt ccagactgag 5880 tgtcagtatgtctacctaca tcagtgtgta agagatgtcc tcagagcaag aaagctacgg 5940 agtgaacaagaaaacccctt gtttccaatc tatgaaaatg tgaatccaga gtatcacaga 6000 gatccagtctattcaaggca ttgagaatgt acctgaagag ctcctggata aaaattattc 6060 actgtgtgatttgtttttaa aaacttgctt catgccctac agaggtgcca gctatttctg 6120 ttgatactatgtataattta ttaatctgga gaatgtttaa aattttatat aatttaaagg 6180 taacagatattattgtacat agttgtattt tgtagtttct tctgtaaata tgtatttttc 6240 ataatgtttaatattaagct ttatataata ctatttttcc acactaaagt gttcatgact 6300 tgttctacataaaactaatt caacctgtat gacaggacta ctggtaaaat gcatatggag 6360 gtggtggcagagacaatcct tcaggccatg ttttctacct gttttgatat tcactggaca 6420 ggaaaaagggcagggctaga gagagcaaat actatggcta gatttgctgc attgctgtcc 6480 catgaatctcgagagccaac agacatgtcc taacttgcta ttaggacaaa tgtgacagtc 6540 aaaaaaaggattagagggag ggagaaaaaa gaattaagca gtaccaaagc tgaactagat 6600 gcttgtgtctgaactagttg ctctctctct cctttctcct tccagggatt caagccaaag 6660 tggtcagctcagggatcatg taacttgcag tgcaagccca ggatggtagg atgcagggtt 6720 gagggttctgatagagaatg attccaaaca gaagtgatga attccttttg ttaataagat 6780 gccagctatacccagactgg aaacataaca tgcaaagcac tatctacagt gattagagat 6840 cctttcattgcattcatggt gtggagtgtg aacatccaca cccatactgt aatgtattta 6900 tacacactagtttctgtctc attttcagtg gtctccattc ctagaaaagt cacaattatc 6960 cattcctacttgatttccca ttaaaagaat attatggtag cagattgtgc ccctcattaa 7020 aaggcttaatgccaacattt tcatagaaat gactacaaac atcatatata gtaaatttaa 7080 aaacaatagcaaaaacaaaa acagtggtct tcagtaaaat tttcaaaact tcttttagta 7140 aatcaatgaagtcaaaatgt caagtaatca cccaaagttg catttaataa caaaaggcac 7200 tacatactgtaccaagttta tcttcaatat ttgtgcctta cttactttga ctataacaaa 7260 ttccaatgagtcagaaagta tttccttcat caaggtccag ttccgacagc attcctggga 7320 aaaatttgaaaggagtttgt acggaatctt catagatacc tgagaagatg agctggagat 7380 gtttgcctttttcacactac aaatttttct gtaataaact tgggaattag aggtcaaaaa 7440 aaaa 7444 382475 DNA Homo sapien misc_feature (1001)..(1001) a, c, g or t 38atcacctgca tcctcgagga cagaccttgt gaagtcagag ctgctacaca ttgaatctca 60agtcgagctt ctgagattcg atgattcagg aagaaaggat tctgaggttt tgaagcaaaa 120tgcagtgaac agcaaccaat ccaatgttgt aattgaagac tttgagtcct cacggatctc 180ttcgtctttg cagcgtagcc cgagtcggtc agcgccggag gacctcagca gccatgtcga 240agccccatag tgaagccggg actgccttca ttcagaccca gcagctgcac gcagccatgg 300ctgacacatt cctggagcac atgtgccgcc tggacattga ttcaccaccc atcacagccc 360ggaacactgg catcatctgt accattgggc ccagcttccc gatcagtgga gacgttgaag 420gagatgataa gtctggaatg aatgtggctc gtctgaattc tctcatggac tcatgagtac 480catgcggaga ccatcaagaa tgtgcgcaca gccacggaaa gctttgcttc tgaccccatc 540ctctaccggc ccgttgctgt ggctctagac actaaaggac ctgagatccg aactgggctc 600atcaagggca gcggcactgc agaggtggag ctgaagaagg gagccactct caaaatcacg 660ctggataacg cctacatgga aaagtgtgac gagaacatcc tgtggctgga ctacaagaac 720atctgcaagg tggtggaagt gggcagcaag atctacgtgg atgatgggct tatttctctc 780caggtgaagc agaaaggtgc cgacttcctg gtgacggagg tggaaaatgg tggctccttg 840ggcagcaaga agggtgtgaa ccttcctggg gctgctgtgg acttgcctgc tgtgtcggag 900aaggacatcc caggatctga aagtttgggg gtcgagcagg atgttgatat ggtgtttggc 960gtcattccat cccgcaaagg catctggatg tcccatggaa ngtttaggaa nggtcctggg 1020gagagaaggg aaaagaaaca tccaagatta tccagcaaaa tcgagaatca tgagggggtt 1080cggaggtttg atgaaatcct ggaggccagt gatgggatca tggtggctcg tggtgatcta 1140ggcattgaga ttcctgcaga gaaggtcttc ttgctcagaa gatgatgatt ggacggtgca 1200acccgagact gggaagcctg tcatctgtgc tactccagat gctggagagc atcgatcaag 1260aagccccgcc ccactcgggc tgaaggcagt gatgtggcca atgcagtcct ggatggagcc 1320gactgcatca tgctgtctgg agaaacagcc aaaggggact atcctctgga ggctgtgcgc 1380atgcagcacc tgattgcccg tgaggcagag gctgccatct accacttgca attatttgag 1440gaactccgcc gcctggcgcc cattaccagc gaccccacag aagccaccgc cgtgggtgcc 1500gtggaggcct ccttcaagtg ctgcagtggg gccataatcg tcctcaccaa gtctggcagg 1560tctgctcacc aggtggccag ataccgccca cgtgccccca tcattgctgt gacccggaat 1620ccccagacag ctcgtcaggc ccacctgtac cgtggcatct tccctgtgct gtgcaaggac 1680ccagtccagg aggcctgggc tgaggacgtg gacctccggg tgaactttgc catgaatgtt 1740ggcaaggccc gaggcttctt caagaaggga gatgtggtca ttgtgctgac cggatggcgc 1800cctggctccg gcttcaccaa caccatgcgt gttgttcctg tgccgtgatg gaccccagag 1860cccctcctcc agcccctgtc ccaccccctt cccccagccc atccattagg ccagcaacgc 1920ttgtagaact cactctgggc tgtaacgtgg cactggtagg ttgggacacc agggaagaag 1980atcaacgcct cactgaaaca tggctgtgtt tgcagcctgc tctagtggga cagcccagag 2040cctggctgcc catcatgtgg ccccacccaa tcaagggaag aaggaggaat gctggactgg 2100aggcccctgg agccagatgg caagagggtg acagcttcct ttcctgtgtg tactctgtcc 2160agttccttta gaaaaaatgg atgcccagag gactcccaac cctggcttgg ggtcaagaaa 2220cagccagcaa gagttagggg ccttagggca ctgggctgtt gttccattga agccgactct 2280ggccctggcc cttacttgct tctctagctc tctaggcctc tccagtttgc acctgtcccc 2340accctccact cagctgtcct gcagcaaaca ctccaccctc caccttccat tttcccccac 2400tactgcagca cctccaggcc tgttgctata gagcctacct gtatgtcaat aaacaacagc 2460tgaagcacca aaaaa 2475 39 298 PRT Homo sapien 39 Trp Ser Tyr Arg Gly GlyGly Arg Tyr Tyr Ala Asp Ser Val Arg Gly 1 5 10 15 Arg Thr Val Ser ArgAsp Asn Ala Lys Asn Ser Tyr Met Asn Ser Arg 20 25 30 Ala Asp Thr Ala ValTyr Cys Ala Arg Ala Asn Tyr Asp Cys Trp Ser 35 40 45 Gly Thr Val Thr ValSer Ser Ala Ser Thr Lys Gly Ser Val Ala Cys 50 55 60 Ser Arg Ser Thr SerGly Gly Thr Ala Ala Gly Cys Val Lys Asp Tyr 65 70 75 80 Val Thr Val SerTrp Asn Ser Gly Ala Thr Ser Gly Val His Thr Ala 85 90 95 Val Ser Ser GlyTyr Ser Ser Ser Val Val Thr Val Ser Ser Ser Gly 100 105 110 Thr Thr TyrThr Cys Asn Val Asn His Lys Ser Asn Thr Lys Val Asp 115 120 125 Lys ArgVal Lys Thr Gly Asp Thr Thr His Thr Cys Arg Cys Lys Ser 130 135 140 CysAsp Thr Cys Arg Cys Lys Ser Cys Asp Thr Cys Arg Cys Ala Gly 145 150 155160 Gly Ser Val Lys Lys Asp Thr Met Ser Arg Thr Val Thr Cys Val Val 165170 175 Val Asp Val Ser His Asp Val Lys Trp Tyr Val Asp Gly Val Val His180 185 190 Asn Ala Lys Thr Lys Arg Asn Ser Thr Arg Val Val Ser Val ThrVal 195 200 205 His Asp Trp Asn Gly Lys Tyr Lys Cys Lys Val Ser Asn LysAla Ala 210 215 220 Lys Thr Ser Lys Thr Lys Gly Arg Val Tyr Thr Ser ArgMet Thr Lys 225 230 235 240 Asn Val Ser Thr Cys Val Lys Gly Tyr Ser AspAla Val Trp Ser Ser 245 250 255 Gly Asn Asn Tyr Asn Thr Thr Met Asp SerAsp Gly Ser Tyr Ser Lys 260 265 270 Thr Val Asp Lys Ser Arg Trp Gly AsnSer Cys Ser Val Met His Ala 275 280 285 His Asn Arg Thr Lys Ser Ser SerGly Lys 290 295 40 168 PRT Homo sapien 40 Pro Gln Leu Ala Cys Leu PheGln Val Lys Ser Gly Ser Pro Ala Val 1 5 10 15 Leu Ala Phe Ala Lys GluLys Ser Phe Gly Trp Pro Ser Phe Ile Thr 20 25 30 Tyr Thr Val Gly Val SerAsp Pro Ala Ala Gly Ser Gln Gly Pro Leu 35 40 45 Ser Thr Thr Leu Thr PheSer Ser Pro Val Thr Asn Gln Ala Ile Ala 50 55 60 Ile Pro Val Thr Val AlaPhe Val Met Asp Arg Arg Gly Pro Gly Pro 65 70 75 80 Tyr Gly Ala Ser LeuPhe Gln His Phe Leu Asp Ser Tyr Gln Val Met 85 90 95 Phe Phe Thr Leu PheAla Leu Leu Ala Gly Thr Ala Val Met Ile Ile 100 105 110 Ala Tyr His ThrVal Cys Thr Pro Arg Asp Leu Ala Val Pro Ala Ala 115 120 125 Leu Thr ProArg Ala Ser Pro Gly His Ser Pro His Tyr Phe Ala Ala 130 135 140 Ser SerPro Thr Ser Pro Asn Ala Leu Pro Pro Ala Arg Lys Ala Ser 145 150 155 160Pro Pro Ser Gly Leu Trp Ser Pro 165 41 78 PRT Homo sapien 41 Val Ser GluGly Ala Thr Trp Ala Ile Gly Phe Pro Ala Ser Phe Pro 1 5 10 15 Leu PheLeu Ala Pro Ala Ala Glu Ala Gly Arg Pro Trp Arg Thr Ser 20 25 30 Trp GlyLeu Thr Ala Ala Ser Pro Gly Ser Ser Trp Gly His Leu Ser 35 40 45 Ser LysVal Cys Thr Gln Glu Val Pro His His Ile Gln Pro His Gly 50 55 60 Ser ProArg Ser Ala Arg Gln Gln Ile Arg Ala Pro Cys His 65 70 75 42 1118 PRTHomo sapien 42 Met Ala Arg Ser Pro Gly Arg Ala Tyr Ala Leu Leu Leu LeuLeu Ile 1 5 10 15 Cys Phe Asn Val Gly Ser Gly Leu His Leu Gln Val LeuSer Thr Arg 20 25 30 Asn Glu Asn Lys Leu Leu Pro Lys His Pro His Leu ValArg Gln Lys 35 40 45 Arg Ala Trp Ile Thr Ala Pro Val Ala Leu Arg Glu GlyGlu Asp Leu 50 55 60 Ser Lys Lys Asn Pro Ile Ala Lys Ile His Ser Asp LeuAla Glu Glu 65 70 75 80 Arg Gly Leu Lys Ile Thr Tyr Lys Tyr Thr Gly LysGly Ile Thr Glu 85 90 95 Pro Pro Phe Gly Ile Phe Val Phe Asn Lys Asp ThrGly Glu Leu Asn 100 105 110 Val Thr Ser Ile Leu Asp Arg Glu Glu Thr ProPhe Phe Leu Leu Thr 115 120 125 Gly Tyr Ala Leu Asp Ala Arg Gly Asn AsnVal Glu Lys Pro Leu Glu 130 135 140 Leu Arg Ile Lys Val Leu Asp Ile AsnAsp Asn Glu Pro Val Phe Thr 145 150 155 160 Gln Asp Val Phe Val Gly SerVal Glu Glu Leu Ser Ala Ala His Thr 165 170 175 Leu Val Met Lys Ile AsnAla Thr Asp Ala Asp Glu Pro Asn Thr Leu 180 185 190 Asn Ser Lys Ile SerTyr Arg Ile Val Ser Leu Glu Pro Ala Tyr Pro 195 200 205 Pro Val Phe TyrLeu Asn Lys Asp Thr Gly Glu Ile Tyr Thr Thr Ser 210 215 220 Val Thr LeuAsp Arg Glu Glu His Ser Ser Tyr Thr Leu Thr Val Glu 225 230 235 240 AlaArg Asp Gly Asn Gly Glu Val Thr Asp Lys Pro Val Lys Gln Ala 245 250 255Gln Val Gln Ile Arg Ile Leu Asp Val Asn Asp Asn Ile Pro Val Val 260 265270 Glu Asn Lys Val Leu Glu Gly Met Val Glu Glu Asn Gln Val Asn Val 275280 285 Glu Val Thr Arg Ile Lys Val Phe Asp Ala Asp Glu Ile Gly Ser Asp290 295 300 Asn Trp Leu Ala Asn Phe Thr Phe Ala Ser Gly Asn Glu Gly GlyTyr 305 310 315 320 Phe His Ile Glu Thr Asp Ala Gln Thr Asn Glu Gly IleVal Thr Leu 325 330 335 Ile Lys Glu Val Asp Tyr Glu Glu Met Lys Asn LeuAsp Phe Ser Val 340 345 350 Ile Val Ala Asn Lys Ala Ala Phe His Lys SerIle Arg Ser Lys Tyr 355 360 365 Lys Pro Thr Pro Ile Pro Ile Lys Val LysVal Lys Asn Val Lys Glu 370 375 380 Gly Ile His Phe Lys Ser Ser Val IleSer Ile Tyr Val Ser Glu Ser 385 390 395 400 Met Asp Arg Ser Ser Lys GlyGln Ile Ile Gly Asn Phe Gln Ala Phe 405 410 415 Asp Glu Asp Thr Gly LeuPro Ala His Ala Arg Tyr Val Lys Leu Glu 420 425 430 Asp Arg Asp Asn TrpIle Ser Val Asp Ser Val Thr Ser Glu Ile Lys 435 440 445 Leu Ala Lys LeuPro Asp Phe Glu Ser Arg Tyr Val Gln Asn Gly Thr 450 455 460 Tyr Thr ValLys Ile Val Ala Ile Ser Glu Asp Tyr Pro Arg Lys Thr 465 470 475 480 IleThr Gly Thr Val Leu Ile Asn Val Glu Asp Ile Asn Asp Asn Cys 485 490 495Pro Thr Leu Ile Glu Pro Val Gln Thr Ile Cys His Asp Ala Glu Tyr 500 505510 Val Asn Val Thr Ala Glu Asp Leu Asp Gly His Pro Asn Ser Gly Pro 515520 525 Phe Ser Phe Ser Val Ile Asp Lys Pro Pro Gly Met Ala Glu Lys Trp530 535 540 Lys Ile Ala Arg Gln Glu Ser Thr Ser Val Leu Leu Gln Gln SerGlu 545 550 555 560 Lys Lys Leu Gly Arg Ser Glu Ile Gln Phe Leu Ile SerAsp Asn Gln 565 570 575 Gly Phe Ser Cys Pro Glu Lys Gln Val Leu Thr LeuThr Val Cys Glu 580 585 590 Cys Leu His Gly Ser Gly Cys Arg Glu Ala GlnHis Asp Ser Tyr Val 595 600 605 Gly Leu Gly Pro Ala Ala Ile Ala Leu MetIle Leu Ala Phe Leu Leu 610 615 620 Leu Leu Leu Val Pro Leu Leu Leu LeuMet Cys His Cys Gly Lys Gly 625 630 635 640 Ala Lys Gly Phe Thr Pro IlePro Gly Thr Ile Glu Met Leu His Pro 645 650 655 Trp Asn Asn Glu Gly AlaPro Pro Glu Asp Lys Val Val Pro Ser Phe 660 665 670 Leu Pro Val Asp GlnGly Gly Ser Leu Val Gly Arg Asn Gly Val Gly 675 680 685 Gly Met Ala LysGlu Ala Thr Met Lys Gly Ser Ser Ser Ala Ser Ile 690 695 700 Val Lys GlyGln His Glu Met Ser Glu Met Asp Gly Arg Trp Glu Glu 705 710 715 720 HisArg Ser Leu Leu Ser Gly Arg Ala Thr Gln Phe Thr Gly Ala Thr 725 730 735Gly Ala Ile Met Thr Thr Glu Thr Thr Lys Thr Ala Arg Ala Thr Gly 740 745750 Ala Ser Arg Asp Met Ala Gly Ala Gln Ala Ala Ala Val Ala Leu Asn 755760 765 Glu Glu Phe Leu Arg Asn Tyr Phe Thr Asp Lys Ala Ala Ser Tyr Thr770 775 780 Glu Glu Asp Glu Asn His Thr Ala Lys Asp Cys Leu Leu Val TyrSer 785 790 795 800 Gln Glu Glu Thr Glu Ser Leu Asn Ala Ser Ile Gly CysCys Ser Phe 805 810 815 Ile Glu Gly Glu Leu Asp Asp Arg Phe Leu Asp AspLeu Gly Leu Lys 820 825 830 Phe Lys Thr Leu Ala Glu Val Cys Leu Gly GlnLys Ile Asp Ile Asn 835 840 845 Lys Glu Ile Glu Gln Arg Gln Lys Pro AlaThr Glu Thr Ser Met Asn 850 855 860 Thr Ala Ser His Ser Leu Cys Glu GlnThr Met Val Asn Ser Glu Asn 865 870 875 880 Thr Tyr Ser Ser Gly Ser SerPhe Pro Val Pro Lys Ser Leu Gln Glu 885 890 895 Ala Asn Ala Glu Lys ValThr Gln Glu Ile Val Thr Glu Arg Ser Val 900 905 910 Ser Ser Arg Gln AlaGln Lys Val Ala Thr Pro Leu Pro Asp Pro Met 915 920 925 Ala Ser Arg AsnVal Ile Ala Thr Glu Thr Ser Tyr Val Thr Gly Ser 930 935 940 Thr Met ProPro Thr Thr Val Ile Leu Gly Pro Ser Gln Pro Gln Ser 945 950 955 960 LeuIle Val Thr Glu Arg Val Tyr Ala Pro Ala Ser Thr Leu Val Asp 965 970 975Gln Pro Tyr Ala Asn Glu Gly Thr Val Val Val Thr Glu Arg Val Ile 980 985990 Gln Pro His Gly Gly Gly Ser Asn Pro Leu Glu Gly Thr Gln His Leu 9951000 1005 Gln Asp Val Pro Tyr Val Met Val Arg Glu Arg Glu Ser Phe Leu1010 1015 1020 Ala Pro Ser Ser Gly Val Gln Pro Thr Leu Ala Met Pro AsnIle 1025 1030 1035 Ala Val Gly Gln Asn Val Thr Val Thr Glu Arg Val LeuAla Pro 1040 1045 1050 Ala Ser Thr Leu Gln Ser Ser Tyr Gln Ile Pro ThrGlu Asn Ser 1055 1060 1065 Met Thr Ala Arg Asn Thr Thr Val Ser Gly AlaGly Val Pro Gly 1070 1075 1080 Pro Leu Pro Asp Phe Gly Leu Glu Glu SerGly His Ser Asn Ser 1085 1090 1095 Thr Ile Thr Thr Ser Ser Thr Arg ValThr Lys His Ser Thr Val 1100 1105 1110 Gln His Ser Tyr Ser 1115 43 97PRT Homo sapien 43 Met Thr Lys Gly Thr Ser Ser Phe Gly Lys Arg Arg AsnLys Thr His 1 5 10 15 Thr Leu Cys Arg Arg Cys Gly Ser Lys Ala Tyr HisLeu Gln Lys Ser 20 25 30 Thr Cys Gly Lys Cys Gly Tyr Pro Ala Lys Arg LysArg Lys Tyr Asn 35 40 45 Trp Ser Ala Lys Ala Lys Arg Arg Asn Thr Thr GlyThr Gly Arg Met 50 55 60 Arg His Leu Lys Ile Val Tyr Arg Arg Phe Arg HisGly Phe Arg Glu 65 70 75 80 Gly Thr Thr Pro Lys Pro Lys Arg Ala Ala ValAla Ala Ser Ser Ser 85 90 95 Ser 44 889 PRT Homo sapien 44 Met Ala AlaAla Val Gly Val Arg Gly Arg Tyr Glu Leu Pro Pro Cys 1 5 10 15 Ser GlyPro Gly Trp Leu Leu Ser Leu Ser Ala Leu Leu Ser Val Ala 20 25 30 Ala ArgGly Ala Phe Ala Thr Thr His Trp Val Val Thr Glu Asp Gly 35 40 45 Lys IleGln Gln Gln Val Asp Ser Pro Met Asn Leu Lys His Pro His 50 55 60 Asp LeuVal Ile Leu Met Arg Gln Glu Ala Thr Val Asn Tyr Leu Lys 65 70 75 80 GluLeu Glu Lys Gln Leu Val Ala Gln Lys Ile His Ile Glu Glu Asn 85 90 95 GluAsp Arg Asp Thr Gly Leu Glu Gln Arg His Asn Lys Glu Asp Pro 100 105 110Asp Cys Ile Lys Ala Lys Val Pro Leu Gly Asp Leu Asp Leu Tyr Asp 115 120125 Gly Thr Tyr Ile Thr Leu Glu Ser Lys Asp Ile Ser Pro Glu Asp Tyr 130135 140 Ile Asp Thr Glu Ser Pro Val Pro Pro Asp Pro Glu Gln Pro Asp Cys145 150 155 160 Thr Lys Ile Leu Glu Leu Pro Tyr Ser Ile His Ala Phe GlnHis Leu 165 170 175 Arg Gly Val Gln Glu Arg Val Asn Leu Ser Ala Pro LeuLeu Pro Lys 180 185 190 Glu Asp Pro Ile Phe Thr Tyr Leu Ser Lys Arg LeuGly Arg Ser Ile 195 200 205 Asp Asp Ile Gly His Leu Ile His Glu Gly LeuGln Lys Asn Thr Ser 210 215 220 Ser Trp Val Leu Tyr Asn Met Ala Ser PheTyr Trp Arg Ile Lys Asn 225 230 235 240 Glu Pro Tyr Gln Val Val Glu CysAla Met Arg Ala Leu His Phe Ser 245 250 255 Ser Arg His Asn Lys Asp IleAla Leu Val Asn Leu Ala Asn Val Leu 260 265 270 His Arg Ala His Phe SerAla Asp Ala Ala Val Val Val His Ala Ala 275 280 285 Leu Asp Asp Ser AspPhe Phe Thr Ser Tyr Tyr Thr Leu Gly Asn Ile 290 295 300 Tyr Ala Met LeuGly Glu Tyr Asn His Ser Val Leu Cys Tyr Asp His 305 310 315 320 Ala LeuGln Ala Arg Pro Gly Phe Glu Gln Ala Ile Lys Arg Lys His 325 330 335 AlaVal Leu Cys Gln Gln Lys Leu Glu Gln Lys Leu Glu Ala Gln His 340 345 350Arg Ser Leu Gln Arg Thr Leu Asn Glu Leu Lys Glu Tyr Gln Lys Gln 355 360365 His Asp His Tyr Leu Arg Gln Gln Glu Ile Leu Glu Lys His Lys Leu 370375 380 Ile Gln Glu Glu Gln Ile Leu Arg Asn Ile Ile His Glu Thr Gln Met385 390 395 400 Ala Lys Glu Ala Gln Leu Gly Asn His Gln Ile Cys Arg LeuVal Asn 405 410 415 Gln Gln His Ser Leu His Cys Gln Trp Asp Gln Pro ValArg Tyr His 420 425 430 Arg Gly Asp Ile Phe Glu Asn Val Asp Tyr Val GlnPhe Gly Glu Asp 435 440 445 Ser Ser Thr Ser Ser Met Met Ser Val Asn PheAsp Val Gln Ser Asn 450 455 460 Gln Ser Asp Ile Asn Asp Ser Val Lys SerSer Pro Val Ala His Ser 465 470 475 480 Ile Leu Trp Ile Trp Gly Arg AspSer Asp Ala Tyr Arg Asp Lys Gln 485 490 495 His Ile Leu Trp Pro Lys ArgAla Asp Cys Thr Glu Ser Tyr Pro Arg 500 505 510 Val Pro Val Gly Gly GluLeu Pro Thr Tyr Phe Leu Pro Pro Glu Asn 515 520 525 Lys Gly Leu Arg IleHis Glu Leu Ser Ser Asp Asp Tyr Ser Thr Glu 530 535 540 Glu Glu Ala GlnThr Pro Asp Cys Ser Ile Thr Asp Phe Arg Lys Ser 545 550 555 560 His ThrLeu Ser Tyr Leu Val Lys Glu Leu Glu Val Arg Met Asp Leu 565 570 575 LysAla Lys Met Pro Asp Asp His Ala Arg Lys Ile Leu Leu Ser Arg 580 585 590Ile Asn Asn Tyr Thr Ile Pro Glu Glu Glu Ile Gly Ser Phe Leu Phe 595 600605 His Ala Ile Asn Lys Pro Asn Ala Pro Ile Trp Leu Ile Leu Asn Glu 610615 620 Ala Gly Leu Tyr Trp Arg Ala Val Gly Asn Ser Thr Phe Ala Ile Ala625 630 635 640 Cys Leu Gln Arg Ala Leu Asn Leu Ala Pro Leu Gln Tyr GlnAsp Val 645 650 655 Pro Leu Val Asn Leu Ala Asn Leu Leu Ile His Tyr GlyLeu His Leu 660 665 670 Asp Ala Thr Lys Leu Leu Leu Gln Ala Leu Ala IleAsn Ser Ser Glu 675 680 685 Pro Leu Thr Phe Leu Ser Leu Gly Asn Ala TyrLeu Ala Leu Lys Asn 690 695 700 Ile Ser Gly Ala Leu Glu Ala Phe Arg GlnAla Leu Lys Leu Thr Thr 705 710 715 720 Lys Cys Pro Glu Cys Glu Asn SerLeu Lys Leu Ile Arg Cys Met Gln 725 730 735 Phe Tyr Pro Phe Leu Tyr AsnIle Thr Ser Ser Val Cys Ser Gly Thr 740 745 750 Val Val Glu Glu Ser AsnGly Ser Asp Glu Met Glu Asn Ser Asp Glu 755 760 765 Thr Lys Met Ser GluGlu Ile Leu Ala Leu Val Asp Glu Phe Gln Gln 770 775 780 Ala Trp Pro LeuGlu Gly Phe Gly Gly Ala Leu Glu Met Lys Gly Arg 785 790 795 800 Arg LeuAsp Leu Gln Gly Ile Arg Val Leu Lys Lys Gly Pro Gln Asp 805 810 815 GlyVal Ala Arg Ser Ser Cys Tyr Gly Asp Cys Arg Ser Glu Asp Asp 820 825 830Glu Ala Thr Glu Trp Ile Thr Phe Gln Val Lys Arg Val Lys Lys Pro 835 840845 Lys Gly Asp His Lys Lys Thr Pro Gly Lys Lys Val Glu Thr Gly Gln 850855 860 Ile Glu Asn Gly His Arg Tyr Gln Ala Asn Leu Glu Ile Thr Gly Pro865 870 875 880 Lys Val Ala Ser Pro Gly Pro Gln Gly 885 45 690 PRT Homosapien 45 Phe Leu Thr Leu Phe Ile Phe Arg Ser Gly Leu Cys Arg Gly AsnSer 1 5 10 15 Val Glu Arg Lys Ile Tyr Ile Pro Leu Asn Lys Thr Ala ProCys Val 20 25 30 Arg Leu Leu Asn Ala Thr His Gln Ile Gly Cys Gln Ser SerIle Ser 35 40 45 Gly Asp Thr Gly Val Ile His Val Val Glu Lys Glu Glu AspLeu Gln 50 55 60 Trp Val Leu Thr Asp Gly Pro Asn Pro Pro Tyr Met Val LeuLeu Glu 65 70 75 80 Ser Lys His Phe Thr Arg Asp Leu Met Glu Lys Leu LysGly Arg Thr 85 90 95 Ser Arg Ile Ala Gly Leu Ala Val Ser Leu Thr Lys ProSer Pro Ala 100 105 110 Ser Gly Phe Ser Pro Ser Val Gln Cys Pro Asn AspGly Phe Gly Val 115 120 125 Tyr Ser Asn Ser Tyr Gly Pro Glu Phe Ala HisCys Arg Glu Ile Gln 130 135 140 Trp Asn Ser Leu Gly Asn Gly Leu Ala TyrGlu Asp Phe Ser Phe Pro 145 150 155 160 Ile Phe Leu Leu Glu Asp Glu AsnGlu Thr Lys Val Ile Lys Gln Cys 165 170 175 Tyr Gln Asp His Asn Leu SerGln Asn Gly Ser Ala Pro Thr Phe Pro 180 185 190 Leu Cys Ala Met Gln LeuPhe Ser His Met His Ala Val Ile Ser Thr 195 200 205 Ala Thr Cys Met ArgArg Ser Ser Ile Gln Ser Thr Phe Ser Ile Asn 210 215 220 Pro Glu Ile ValCys Asp Pro Leu Ser Asp Tyr Asn Val Trp Ser Met 225 230 235 240 Leu LysPro Ile Asn Thr Thr Gly Thr Leu Lys Pro Asp Asp Arg Val 245 250 255 ValVal Ala Ala Thr Arg Leu Asp Ser Arg Ser Phe Phe Trp Asn Val 260 265 270Ala Pro Gly Ala Glu Ser Ala Val Ala Ser Phe Val Thr Gln Leu Ala 275 280285 Ala Ala Glu Ala Leu Gln Lys Ala Pro Asp Val Thr Thr Leu Pro Arg 290295 300 Asn Val Met Phe Val Phe Phe Gln Gly Glu Thr Phe Asp Tyr Ile Gly305 310 315 320 Ser Ser Arg Met Val Tyr Asp Met Glu Lys Gly Lys Phe ProVal Gln 325 330 335 Leu Glu Asn Val Asp Ser Phe Val Glu Leu Gly Gln ValAla Leu Arg 340 345 350 Thr Ser Leu Glu Leu Trp Met His Thr Asp Pro ValSer Gln Lys Asn 355 360 365 Glu Ser Val Arg Asn Gln Val Glu Asp Leu LeuAla Thr Leu Glu Lys 370 375 380 Ser Gly Ala Gly Val Pro Ala Val Ile LeuArg Arg Pro Asn Gln Ser 385 390 395 400 Gln Pro Leu Pro Pro Ser Ser LeuGln Arg Phe Leu Arg Ala Arg Asn 405 410 415 Ile Ser Gly Val Val Leu AlaAsp His Ser Gly Ala Phe His Asn Lys 420 425 430 Tyr Tyr Gln Ser Ile TyrAsp Thr Ala Glu Asn Ile Asn Val Ser Tyr 435 440 445 Pro Glu Trp Leu SerPro Glu Glu Asp Leu Asn Phe Val Thr Asp Thr 450 455 460 Ala Lys Ala LeuAla Asp Val Ala Thr Val Leu Gly Arg Ala Leu Tyr 465 470 475 480 Glu LeuAla Gly Gly Thr Asn Phe Ser Asp Thr Val Gln Ala Asp Pro 485 490 495 GlnThr Val Thr Arg Leu Leu Tyr Gly Phe Leu Ile Lys Ala Asn Asn 500 505 510Ser Trp Phe Gln Ser Ile Leu Arg Gln Asp Leu Arg Ser Tyr Leu Gly 515 520525 Asp Gly Pro Leu Gln His Tyr Ile Ala Val Ser Ser Pro Thr Asn Thr 530535 540 Thr Tyr Val Val Gln Tyr Ala Leu Ala Asn Leu Thr Gly Thr Val Val545 550 555 560 Asn Leu Thr Arg Glu Gln Cys Gln Asp Pro Ser Lys Val ProSer Glu 565 570 575 Asn Lys Asp Leu Tyr Glu Tyr Ser Trp Val Gln Gly ProLeu His Ser 580 585 590 Asn Glu Thr Asp Arg Leu Pro Arg Cys Val Arg SerThr Ala Arg Leu 595 600 605 Ala Arg Ala Leu Ser Pro Ala Phe Glu Leu SerGln Trp Ser Ser Thr 610 615 620 Glu Tyr Ser Thr Trp Thr Glu Ser Arg TrpLys Asp Ile Arg Ala Arg 625 630 635 640 Ile Phe Leu Ile Ala Ser Lys GluLeu Glu Leu Ile Thr Leu Thr Val 645 650 655 Gly Phe Gly Ile Leu Ile PheSer Leu Ile Val Thr Tyr Cys Ile Asn 660 665 670 Ala Lys Ala Asp Val LeuPhe Ile Ala Pro Arg Glu Pro Gly Ala Val 675 680 685 Ser Tyr 690 46 170PRT Homo sapien 46 Gln Val Pro Arg Ser Lys Ala Leu Glu Val Thr Lys LeuAla Ile Glu 1 5 10 15 Ala Gly Phe Arg His Ile Asp Ser Ala His Leu TyrAsn Asn Glu Glu 20 25 30 Gln Val Gly Leu Ala Ile Arg Ser Lys Ile Ala AspGly Ser Val Lys 35 40 45 Arg Glu Asp Ile Phe Tyr Thr Ser Lys Leu Trp SerThr Phe His Arg 50 55 60 Pro Glu Leu Val Arg Pro Ala Leu Glu Asn Ser LeuLys Lys Ala Gln 65 70 75 80 Leu Asp Tyr Val Asp Leu Tyr Leu Ile His SerPro Met Ser Leu Lys 85 90 95 Pro Gly Glu Glu Leu Ser Pro Thr Asp Glu GlnVal Ala Lys Val Ile 100 105 110 Phe Asp Ile Val Asp Leu Cys Thr Thr TrpGlu Gly Met Glu Lys Cys 115 120 125 Lys Asp Gly Arg Asn Trp Gly Lys SerIle Gly Val Ser His Phe Asn 130 135 140 Pro Gln Ala Leu Gly Met Ser LeuGln Lys Ala Gly Ile Gln Leu Lys 145 150 155 160 Arg Ser Ala Pro Val GluCys Pro Ile Tyr 165 170 47 1596 PRT Homo sapien 47 Met Thr Thr Glu ThrGly Pro Asp Ser Glu Val Lys Lys Ala Gln Glu 1 5 10 15 Glu Ala Pro GlnGln Pro Glu Ala Ala Ala Ala Val Thr Thr Pro Val 20 25 30 Thr Pro Ala GlyHis Gly His Pro Glu Ala Asn Ser Asn Glu Lys His 35 40 45 Pro Ser Gln GlnAsp Thr Arg Pro Ala Glu Gln Ser Leu Asp Met Glu 50 55 60 Glu Lys Asp TyrSer Glu Ala Asp Gly Leu Ser Glu Arg Thr Thr Pro 65 70 75 80 Ser Lys AlaGln Lys Ser Pro Gln Lys Ile Ala Lys Lys Tyr Lys Ser 85 90 95 Ala Ile CysArg Val Thr Leu Leu Asp Ala Ser Glu Tyr Glu Cys Glu 100 105 110 Val GluLys His Gly Arg Gly Gln Val Leu Phe Asp Leu Val Cys Glu 115 120 125 HisLeu Asn Leu Leu Glu Lys Asp Tyr Phe Gly Leu Thr Phe Cys Asp 130 135 140Ala Asp Ser Gln Lys Asn Trp Leu Asp Pro Ser Lys Glu Ile Lys Lys 145 150155 160 Gln Ile Arg Ser Glu Trp Leu Val Val Phe Gly Glu Val Gly Ser Pro165 170 175 Trp Asn Phe Ala Phe Thr Val Lys Phe Tyr Pro Pro Asp Pro AlaGln 180 185 190 Leu Thr Glu Asp Ile Thr Arg Tyr Tyr Leu Cys Leu Gln LeuArg Ala 195 200 205 Asp Ile Ile Thr Gly Arg Leu Pro Cys Ser Phe Val ThrHis Ala Leu 210 215 220 Leu Gly Ser Tyr Ala Val Gln Ala Glu Leu Gly AspTyr Asp Ala Glu 225 230 235 240 Glu His Val Gly Asn Tyr Val Ser Glu LeuArg Phe Ala Pro Asn Gln 245 250 255 Thr Arg Glu Leu Glu Glu Arg Ile MetGlu Leu His Lys Thr Tyr Arg 260 265 270 Gly Met Thr Pro Gly Glu Ala GluIle His Phe Leu Glu Asn Ala Lys 275 280 285 Lys Leu Ser Met Tyr Gly ValAsp Leu His His Ala Lys Asp Ser Glu 290 295 300 Gly Ile Asp Ile Met LeuGly Val Cys Ala Asn Gly Leu Leu Ile Tyr 305 310 315 320 Arg Asp Arg LeuArg Ile Asn Arg Phe Ala Trp Pro Lys Ile Leu Lys 325 330 335 Ile Ser TyrLys Arg Ser Asn Phe Tyr Ile Lys Ile Arg Pro Gly Glu 340 345 350 Tyr GluGln Phe Glu Ser Thr Ile Gly Phe Lys Leu Pro Asn His Arg 355 360 365 SerAla Lys Arg Leu Trp Lys Val Cys Ile Glu His His Thr Phe Phe 370 375 380Arg Leu Val Ser Pro Glu Pro Pro Pro Lys Gly Phe Leu Val Met Gly 385 390395 400 Ser Lys Phe Arg Tyr Ser Gly Arg Thr Gln Ala Gln Thr Arg Gln Ala405 410 415 Ser Ala Leu Ile Asp Arg Pro Ala Pro Phe Phe Glu Arg Ser SerSer 420 425 430 Lys Arg Tyr Thr Met Ser Arg Ser Leu Asp Gly Ala Glu PheSer Arg 435 440 445 Pro Ala Ser Val Ser Glu Asn His Asp Ala Gly Pro AspGly Asp Lys 450 455 460 Arg Asp Glu Asp Gly Glu Ser Gly Gly Gln Arg SerGlu Ala Glu Glu 465 470 475 480 Gly Glu Val Arg Thr Pro Thr Lys Ile LysGlu Leu Lys Phe Leu Asp 485 490 495 Lys Pro Glu Asp Val Leu Leu Lys HisGln Ala Ser Ile Asn Glu Leu 500 505 510 Lys Arg Thr Leu Lys Glu Pro AsnSer Lys Leu Ile His Arg Asp Arg 515 520 525 Asp Trp Glu Arg Glu Arg ArgLeu Pro Ser Ser Pro Ala Ser Pro Ser 530 535 540 Pro Lys Gly Thr Pro GluLys Ala Asn Glu Ser Gln Arg Thr Gln Asp 545 550 555 560 Ile Ser Gln ArgAsp Leu Val Pro Glu Pro Gly Ala Ala Ala Gly Leu 565 570 575 Glu Val PheThr Gln Lys Ser Leu Ala Ala Ser Pro Glu Gly Ser Glu 580 585 590 His TrpVal Phe Ile Glu Arg Glu Tyr Thr Arg Pro Glu Glu Leu Gly 595 600 605 LeuLeu Lys Val Thr Thr Met Gln Gln Glu Glu Arg Gln Ala Gly Leu 610 615 620Ala Gly Ile Leu Ala Asn Gly Arg Leu Ser Lys Val Asp Val Leu Val 625 630635 640 Asp Lys Phe Lys Val Glu Val Ala Thr Glu Glu Met Val Gly Asn Arg645 650 655 Arg Ala Asn Thr Gln Gln Gln Gly Lys Met Ile Ala Ser Pro GluAsp 660 665 670 Phe Glu Ser Val Gly Glu Glu Gly Pro Trp Ile Arg Glu SerPro Gly 675 680 685 Gly Ala Ala Leu Ala Ser Gly Arg Thr Leu Ala Glu LysLeu Leu Glu 690 695 700 Gly Ser Glu Leu Arg Ala Asp Thr Arg Glu Ala ThrIle Arg Asn Arg 705 710 715 720 Cys Met Ser Asp Gly Gln Pro Glu Gly GlnThr Glu Leu Arg Lys Gly 725 730 735 Leu Glu Glu Pro His Thr Cys Gly ArgPro Thr Ala Pro Gly Thr Arg 740 745 750 Pro Ala Glu Val Asp Val Leu SerPro Ala Ser Asp Lys Gly Gly Leu 755 760 765 Gln Ser Phe Leu Leu Asp ProAla His Ala Glu Ala Arg Ala Glu Leu 770 775 780 Ser Asn Glu Thr Asp ThrSer Phe Ala Glu Arg Ser Phe Tyr Leu Asn 785 790 795 800 Tyr Glu Glu LysAsp Ser Glu Asp Gln Val Leu Pro Pro Pro Leu Glu 805 810 815 Glu Arg LysGly Arg Leu Asp Ala Pro Pro Gly Gly Glu Pro Arg Pro 820 825 830 Thr LeuAsn Ser Leu Asp Leu Arg Val Ser Ala Ala Ala Ser Ser Arg 835 840 845 SerLys Asp Glu Ala His Met Thr Ser Pro Lys Glu Gly Ala Gly Thr 850 855 860Pro Lys Asn His Gly Gly Pro Gly Asp Leu Lys Gly Ser Pro Ala Gly 865 870875 880 Gln Thr Phe Ala Glu Gly Trp Glu Asp Ala Gln Trp Gly Val Glu Gly885 890 895 Glu Phe Pro His Leu Thr Ala Ser Ala Ala Arg Glu Glu Gly ThrPro 900 905 910 Val Ser Gly Asp Leu Leu Gly Lys Ala Glu Glu Ser Pro ThrGlu Glu 915 920 925 Leu Lys Lys His Pro Pro His Arg Gly Gln Gly Val HisPro Asp Pro 930 935 940 Gln Ala Cys Ala Leu Pro Arg Ala Ile Pro Leu AsnVal Arg Lys Pro 945 950 955 960 Val Lys Pro Asp Arg Gly Asn Phe Pro ProLys Glu Arg Gly Val Val 965 970 975 Pro Thr Gln Lys Gly Gly Ala Glu LeuLys Asp Arg Glu Ala Ser Ala 980 985 990 Phe Leu His Met Glu Val Ile IlePro Leu Pro Ala Ser Pro Gly His 995 1000 1005 Ser Glu Asp Leu Ala AlaLeu Glu Glu Ala Ser Pro Ser Pro Thr 1010 1015 1020 Ser His Gly Ser GlyGlu Pro Ser Glu Leu Arg Glu Pro Phe Leu 1025 1030 1035 Arg His Val HisLeu Ser Lys Ala Ser Pro Glu Pro Lys Asp Gln 1040 1045 1050 Val Gly PheVal Val Ser Pro Ala Thr Gly Gly Glu Arg Arg Pro 1055 1060 1065 Pro ProIle Thr Ser Arg Lys Pro Arg Val Val Pro Glu Glu Ala 1070 1075 1080 GluGly Arg Ile Pro Leu Gly Phe Gly Phe Pro Ser Gly Lys Arg 1085 1090 1095Arg Glu Met Thr Ser Phe Gln Ala Gly Asp Gln Glu Gly Ser Leu 1100 11051110 Glu Asp Ile Ser Lys Thr Ser Val Ala Asn Lys Ile Arg Ile Phe 11151120 1125 Glu Thr His Gly Ala Glu Thr Arg Arg Met Ser Glu Gly Glu Ala1130 1135 1140 Arg Ser Leu Pro Asn Asp Val Ser Ser Glu Ala Pro Val GlyGln 1145 1150 1155 Ala Glu Gln Gln Arg Ser Thr Leu Ser Asp Leu Gly PheAla Gln 1160 1165 1170 Leu Gln Pro Pro Gly Asp Phe Ala Ser Pro Lys AlaThr His Ser 1175 1180 1185 Thr Val Ile Pro Leu Ala Thr Arg His Phe ArgGlu Asp Thr Ser 1190 1195 1200 Ala Ser Tyr Gln Glu Ala His Thr Glu LeuGlu Pro Val Ser Pro 1205 1210 1215 Asn Ser Gly Cys Glu Thr Thr Leu AlaGlu Ala Thr Gly Thr Gly 1220 1225 1230 Val Thr Gly Arg Asn Lys Ser GlyAsp Ala Val Arg Glu Glu Lys 1235 1240 1245 Arg Ser Thr Asn Leu Ala AlaAsn Thr Pro Gly Lys Gly Gly Arg 1250 1255 1260 Leu Arg Phe Ala Ser ProSer Gly Pro Gln Arg Ala Gly Leu Arg 1265 1270 1275 Glu Gly Ser Glu GluLys Val Lys Pro Pro Arg Pro Arg Ala Pro 1280 1285 1290 Glu Ser Asp ThrGly Asp Glu Asp Gln Asp Gln Glu Arg Asp Thr 1295 1300 1305 Val Phe LeuLys Asp Asn His Leu Ala Ile Glu Arg Lys Cys Ser 1310 1315 1320 Ser IleThr Val Ser Ser Thr Ser Ser Leu Glu Ala Glu Val Asp 1325 1330 1335 PheThr Val Ile Gly Asp Tyr His Gly Ser Ala Phe Glu Asp Phe 1340 1345 1350Ser Arg Ser Leu Pro Glu Leu Asp Arg Asp Lys Ser Asp Ser Asp 1355 13601365 Thr Glu Gly Leu Leu Phe Ser Arg Asp Leu Asn Lys Gly Ala Pro 13701375 1380 Ser Gln Asp Asp Glu Ser Gly Gly Ile Glu Asp Ser Pro Asp Arg1385 1390 1395 Gly Ala Cys Ser Thr Pro Asp Met Pro Gln Phe Glu Pro ValLys 1400 1405 1410 Thr Glu Thr Met Thr Val Ser Ser Leu Ala Ile Arg LysLys Ile 1415 1420 1425 Glu Pro Glu Ala Val Leu Gln Thr Arg Val Ser AlaMet Asp Asn 1430 1435 1440 Thr Gln Val Asp Gly Ser Ala Ser Val Gly ArgGlu Phe Ile Ala 1445 1450 1455 Thr Thr Pro Ser Ile Thr Thr Glu Thr IleSer Thr Thr Met Glu 1460 1465 1470 Asn Ser Leu Lys Ser Gly Lys Gly AlaAla Ala Met Ile Pro Gly 1475 1480 1485 Pro Gln Thr Val Ala Thr Glu IleArg Ser Leu Ser Pro Ile Ile 1490 1495 1500 Gly Lys Asp Val Leu Thr SerThr Tyr Gly Ala Thr Ala Glu Thr 1505 1510 1515 Leu Ser Thr Ser Thr ThrThr His Val Thr Lys Thr Val Lys Gly 1520 1525 1530 Gly Phe Ser Glu ThrArg Ile Glu Lys Arg Ile Ile Ile Thr Gly 1535 1540 1545 Asp Glu Asp ValAsp Gln Asp Gln Ala Leu Ala Leu Ala Ile Lys 1550 1555 1560 Glu Ala LysLeu Gln His Pro Asp Met Leu Val Thr Lys Ala Val 1565 1570 1575 Val TyrArg Glu Thr Asp Pro Ser Pro Glu Glu Arg Asp Lys Lys 1580 1585 1590 ProGln Lys 1595 48 455 PRT Homo sapien 48 Met Ala Ala Pro Glu Glu His AspSer Pro Thr Glu Ala Ser Gln Pro 1 5 10 15 Ile Val Glu Glu Glu Glu ThrLys Thr Phe Lys Asp Leu Gly Val Thr 20 25 30 Asp Val Leu Cys Glu Ala CysAsp Gln Leu Gly Trp Thr Lys Pro Thr 35 40 45 Lys Ile Gln Ile Glu Ala IlePro Leu Ala Leu Gln Gly Arg Asp Ile 50 55 60 Ile Gly Leu Ala Glu Thr GlySer Gly Lys Thr Gly Ala Phe Ala Leu 65 70 75 80 Pro Ile Leu Asn Ala LeuLeu Glu Thr Pro Gln Arg Leu Phe Ala Leu 85 90 95 Val Leu Thr Pro Thr ArgGlu Leu Ala Phe Gln Ile Ser Glu Gln Phe 100 105 110 Glu Ala Leu Gly SerSer Ile Gly Val Gln Ser Ala Val Ile Val Gly 115 120 125 Gly Ile Asp SerMet Ser Gln Ser Leu Ala Leu Ala Lys Lys Pro His 130 135 140 Ile Ile IleAla Thr Pro Gly Arg Leu Ile Asp His Leu Glu Asn Thr 145 150 155 160 LysGly Phe Asn Leu Arg Ala Leu Lys Tyr Leu Val Met Asp Glu Ala 165 170 175Asp Arg Ile Leu Asn Met Asp Phe Glu Thr Glu Val Asp Lys Ile Leu 180 185190 Lys Val Ile Pro Arg Asp Arg Lys Thr Phe Leu Phe Ser Ala Thr Met 195200 205 Thr Lys Lys Val Gln Lys Leu Gln Arg Ala Ala Leu Lys Asn Pro Val210 215 220 Lys Cys Ala Val Ser Ser Lys Tyr Gln Thr Val Glu Lys Leu GlnGln 225 230 235 240 Tyr Tyr Ile Phe Ile Pro Ser Lys Phe Lys Asp Thr TyrLeu Val Tyr 245 250 255 Ile Leu Asn Glu Leu Ala Gly Asn Ser Phe Met IlePhe Cys Ser Thr 260 265 270 Cys Asn Asn Thr Gln Arg Thr Ala Leu Leu LeuArg Asn Leu Gly Phe 275 280 285 Thr Ala Ile Pro Leu His Gly Gln Met SerGln Ser Lys Arg Leu Gly 290 295 300 Ser Leu Asn Lys Phe Lys Ala Lys AlaArg Ser Ile Leu Leu Ala Thr 305 310 315 320 Asp Val Ala Ser Arg Gly LeuAsp Ile Pro His Val Asp Val Val Val 325 330 335 Asn Phe Asp Ile Pro ThrHis Ser Lys Asp Tyr Ile His Arg Val Gly 340 345 350 Arg Thr Ala Arg AlaGly Arg Ser Gly Lys Ala Ile Thr Phe Val Thr 355 360 365 Gln Tyr Asp ValGlu Leu Phe Gln Arg Ile Glu His Leu Ile Gly Lys 370 375 380 Lys Leu ProGly Phe Pro Thr Gln Asp Asp Glu Val Met Met Leu Thr 385 390 395 400 GluArg Val Ala Glu Ala Gln Arg Phe Ala Arg Met Glu Leu Arg Glu 405 410 415His Gly Glu Lys Lys Lys Arg Ser Arg Glu Asp Ala Gly Asp Asn Asp 420 425430 Asp Thr Glu Gly Ala Ile Gly Val Arg Asn Lys Val Ala Gly Gly Lys 435440 445 Met Lys Lys Arg Lys Gly Arg 450 455 49 246 PRT Homo sapien 49Met Ala Trp Ala Pro Leu Leu Leu Thr Leu Leu Ser Leu Leu Thr Gly 1 5 1015 Ser Leu Ser Gln Pro Ile Leu Thr Gln Pro Pro Ser Ala Ser Ala Ser 20 2530 Leu Gly Ala Ser Val Thr Leu Thr Cys Ser Val Ser Ser Asp Tyr Lys 35 4045 Asn Leu Glu Val Asp Trp Phe Gln Gln Arg Pro Gly Lys Gly Pro Arg 50 5560 Phe Val Met Arg Val Gly Thr Gly Gly Val Val Gly Phe Arg Gly Ala 65 7075 80 Asp Ile Pro Asp Arg Phe Ser Val Ser Gly Ser Gly Leu Asn Arg Phe 8590 95 Leu Thr Ile Arg Asn Ile Glu Glu Glu Asp Glu Ser Asp Tyr His Cys100 105 110 Gly Thr Asp Leu Gly Ser Gly Thr Ser Phe Val Ser Trp Val PheGly 115 120 125 Gly Gly Thr Lys Leu Thr Val Leu Ser Gln Pro Lys Ala AlaPro Ser 130 135 140 Val Thr Leu Phe Pro Pro Ser Ser Glu Glu Leu Gln AlaAsn Lys Ala 145 150 155 160 Thr Leu Val Cys Leu Ile Ser Asp Phe Tyr ProGly Ala Val Thr Val 165 170 175 Ala Trp Lys Ala Asp Ser Ser Pro Val LysAla Gly Val Glu Thr Thr 180 185 190 Thr Pro Ser Lys Gln Ser Asn Asn LysTyr Ala Ala Ser Ser Tyr Leu 195 200 205 Ser Leu Thr Pro Glu Gln Trp LysSer Asn Arg Ser Tyr Ser Cys Gln 210 215 220 Val Thr His Glu Gly Ser ThrVal Glu Lys Thr Val Ala Pro Thr Glu 225 230 235 240 Cys Ser Thr Glu CysSer 245 50 228 PRT Homo sapien 50 Ala Asn Ala Leu Gly Pro Cys Ala GluIle Val Met Thr Gln Thr Pro 1 5 10 15 Leu Ser Leu Ser Ile Thr Pro GlyGlu Gln Ala Ser Met Ser Cys Arg 20 25 30 Ser Ser Gln Ser Leu Leu His SerAsp Gly Tyr Thr Tyr Leu Tyr Trp 35 40 45 Phe Leu Gln Lys Pro Gly Gln SerPro Gln Leu Leu Ile Tyr Glu Val 50 55 60 Ser Asn Arg Phe Ser Gly Val SerPro Ile Arg Phe Ser Gly Ser Gly 65 70 75 80 Ser Gly Arg Glu Phe Thr LeuArg Ile Ser Arg Val Glu Ala Asp Asp 85 90 95 Ala Gly Val Tyr Tyr Cys MetGln Thr Thr Gln Thr Pro Asn Thr Phe 100 105 110 Gly Gln Gly Thr Arg LeuGlu Ile Lys Arg Thr Val Ala Ala Pro Ser 115 120 125 Val Phe Ile Phe ProPro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala 130 135 140 Ser Val Val CysLeu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val 145 150 155 160 Gln TrpLys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser 165 170 175 ValThr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr 180 185 190Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Leu Tyr Ala Cys 195 200205 Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn 210215 220 Arg Gly Glu Cys 225 51 106 PRT Homo sapien 51 Gly Gln Pro LysAla Asn Pro Thr Val Thr Leu Phe Pro Pro Ser Ser 1 5 10 15 Glu Glu LeuGln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp 20 25 30 Phe Tyr ProGly Ala Val Thr Val Ala Trp Lys Ala Asp Gly Ser Pro 35 40 45 Val Lys AlaGly Val Glu Thr Thr Lys Pro Ser Lys Gln Ser Asn Asn 50 55 60 Lys Tyr AlaAla Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys 65 70 75 80 Ser HisArg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val 85 90 95 Glu LysThr Val Ala Pro Thr Glu Cys Ser 100 105 52 56 PRT Homo sapien 52 Arg ThrGly Tyr Glu Glu Glu Thr Trp Asn Leu Lys Glu Cys Val Gly 1 5 10 15 ArgCys Ala Asn Pro Asn Val Asn Phe Leu Thr Lys Val Glu Ser Pro 20 25 30 GlyMet Val Gln Arg Trp Gly Leu Leu Leu Cys Arg Arg Asp Ser Arg 35 40 45 PheThr Pro Trp Gln Lys Ile Tyr 50 55 53 824 PRT Homo sapien 53 Met Ala PheAla Ser Phe Arg Arg Ile Leu Ala Leu Ser Thr Phe Glu 1 5 10 15 Lys ArgLys Ser Arg Glu Tyr Glu His Val Arg Arg Asp Leu Asp Pro 20 25 30 Asn GluVal Trp Glu Ile Val Gly Glu Leu Gly Asp Gly Ser Phe Gly 35 40 45 Met ValTyr Lys Ala Lys Asn Lys Glu Thr Gly Ala Leu Ala Ala Ala 50 55 60 Ile ValIle Glu Thr Lys Ser Glu Glu Glu Leu Glu Asp Tyr Ile Val 65 70 75 80 GluIle Glu Ile Leu Ala Thr Cys Asp His Pro Tyr Ile Val Lys Leu 85 90 95 LeuGly Ala Tyr Tyr His Asp Gly Lys Leu Trp Ile Met Ile Glu Phe 100 105 110Cys Pro Gly Gly Ala Val Asp Ala Ile Met Leu Glu Leu Asp Arg Gly 115 120125 Leu Thr Glu Pro Gln Ile Gln Val Val Cys Arg Gln Met Leu Glu Ala 130135 140 Leu Asn Phe Leu His Ser Lys Arg Ile Ile His Arg Asp Leu Lys Ala145 150 155 160 Gly Asn Val Leu Met Thr Leu Glu Gly Asp Ile Arg Leu AlaAsp Phe 165 170 175 Gly Val Ser Ala Lys Asn Leu Lys Thr Leu Gln Lys ArgAsp Ser Phe 180 185 190 Ile Gly Thr Pro Tyr Trp Met Ala Pro Glu Val ValMet Cys Glu Thr 195 200 205 Met Lys Asp Thr Pro Tyr Asp Tyr Lys Ala AspIle Trp Ser Leu Gly 210 215 220 Ile Thr Leu Ile Glu Met Ala Gln Ile GluPro Pro His His Glu Leu 225 230 235 240 Asn Pro Met Arg Val Leu Leu LysIle Ala Lys Ser Asp Pro Pro Thr 245 250 255 Leu Leu Thr Pro Ser Lys TrpSer Val Glu Phe Arg Asp Phe Leu Lys 260 265 270 Ile Ala Leu Asp Lys AsnPro Glu Thr Arg Pro Ser Ala Ala Ala Ala 275 280 285 Leu Glu His Pro PheVal Ser Ser Ile Thr Ser Asn Lys Ala Leu Arg 290 295 300 Glu Leu Val AlaGlu Ala Lys Ala Glu Val Met Glu Glu Ile Glu Asp 305 310 315 320 Gly ArgAsp Glu Gly Glu Glu Glu Asp Ala Val Asp Ala Ala Ser Thr 325 330 335 LeuGlu Asn His Thr Gln Asn Ser Ser Glu Val Ser Pro Pro Ser Leu 340 345 350Asn Ala Asp Lys Pro Leu Glu Glu Ser Pro Ser Thr Pro Leu Ala Pro 355 360365 Ser Gln Ser Gln Asp Ser Val Asn Glu Pro Cys Ser Gln Pro Ser Gly 370375 380 Asp Arg Ser Leu Gln Thr Thr Ser Pro Pro Val Val Ala Pro Gly Asn385 390 395 400 Glu Asn Gly Leu Ala Val Pro Val Pro Leu Arg Lys Ser ArgPro Val 405 410 415 Ser Met Asp Ala Arg Ile Gln Val Ala Gln Glu Lys GlnVal Ala Glu 420 425 430 Gln Gly Gly Asp Leu Ser Pro Ala Ala Asn Arg SerGln Lys Ala Ser 435 440 445 Gln Ser Arg Pro Asn Ser Ser Ala Leu Glu ThrLeu Gly Gly Glu Lys 450 455 460 Leu Ala Asn Gly Ser Leu Glu Pro Pro AlaGln Ala Ala Pro Gly Pro 465 470 475 480 Ser Lys Arg Asp Ser Asp Cys SerSer Leu Cys Thr Ser Glu Ser Met 485 490 495 Asp Tyr Gly Thr Asn Leu SerThr Asp Leu Ser Leu Asn Lys Glu Met 500 505 510 Gly Ser Leu Ser Ile LysAsp Pro Lys Leu Tyr Lys Lys Thr Leu Lys 515 520 525 Arg Thr Arg Lys PheVal Val Asp Gly Val Glu Val Ser Ile Thr Thr 530 535 540 Ser Lys Ile IleSer Glu Asp Glu Lys Lys Asp Glu Glu Met Arg Phe 545 550 555 560 Leu ArgArg Gln Glu Leu Arg Glu Leu Arg Leu Leu Gln Lys Glu Glu 565 570 575 HisArg Asn Gln Thr Gln Leu Ser Asn Lys His Glu Leu Gln Leu Glu 580 585 590Gln Met His Lys Arg Phe Glu Gln Glu Ile Asn Ala Lys Lys Lys Phe 595 600605 Phe Asp Thr Glu Leu Glu Asn Leu Glu Arg Gln Gln Lys Gln Gln Val 610615 620 Glu Lys Met Glu Gln Asp His Ala Val Arg Arg Arg Glu Glu Ala Arg625 630 635 640 Arg Ile Arg Leu Glu Gln Asp Arg Asp Tyr Thr Arg Phe GlnGlu Gln 645 650 655 Leu Lys Leu Met Lys Lys Glu Val Lys Asn Glu Val GluLys Leu Pro 660 665 670 Arg Gln Gln Arg Lys Glu Ser Met Lys Gln Lys MetGlu Glu His Thr 675 680 685 Gln Lys Lys Gln Leu Leu Asp Arg Asp Phe ValAla Lys Gln Lys Glu 690 695 700 Asp Leu Glu Leu Ala Met Lys Arg Leu ThrThr Asp Asn Arg Arg Glu 705 710 715 720 Ile Cys Asp Lys Glu Arg Glu CysLeu Met Lys Lys Gln Glu Leu Leu 725 730 735 Arg Asp Arg Glu Ala Ala LeuTrp Glu Met Glu Glu His Gln Leu Gln 740 745 750 Glu Arg His Gln Leu ValLys Gln Gln Leu Lys Asp Gln Tyr Phe Leu 755 760 765 Gln Arg His Glu LeuLeu Arg Lys His Glu Lys Glu Arg Glu Gln Met 770 775 780 Gln Arg Tyr AsnGln Arg Met Ile Glu Gln Leu Lys Val Arg Gln Gln 785 790 795 800 Gln GluLys Ala Arg Leu Pro Lys Ile Gln Arg Ser Glu Gly Lys Thr 805 810 815 ArgMet Ala Met Tyr Lys Lys Ser 820 54 1997 PRT Homo sapien 54 Met Leu SerHis Gly Ala Gly Leu Ala Leu Trp Ile Thr Leu Ser Leu 1 5 10 15 Leu GlnThr Gly Leu Ala Glu Pro Glu Arg Cys Asn Phe Thr Leu Ala 20 25 30 Glu SerLys Ala Ser Ser His Ser Val Ser Ile Gln Trp Arg Ile Leu 35 40 45 Gly SerPro Cys Asn Phe Ser Leu Ile Tyr Ser Ser Asp Thr Leu Gly 50 55 60 Ala AlaLeu Cys Pro Thr Phe Arg Ile Asp Asn Thr Thr Tyr Gly Cys 65 70 75 80 AsnLeu Gln Asp Leu Gln Ala Gly Thr Ile Tyr Asn Phe Arg Ile Ile 85 90 95 SerLeu Asp Glu Glu Arg Thr Val Val Leu Gln Thr Asp Pro Leu Pro 100 105 110Pro Ala Arg Phe Gly Val Ser Lys Glu Lys Thr Thr Ser Thr Ser Leu 115 120125 His Val Trp Trp Thr Pro Ser Ser Gly Lys Val Thr Ser Tyr Glu Val 130135 140 Gln Leu Phe Asp Glu Asn Asn Gln Lys Ile Gln Gly Val Gln Ile Gln145 150 155 160 Glu Ser Thr Ser Trp Asn Glu Tyr Thr Phe Phe Asn Leu ThrAla Gly 165 170 175 Ser Lys Tyr Asn Ile Ala Ile Thr Ala Val Ser Gly GlyLys Arg Ser 180 185 190 Phe Ser Val Tyr Thr Asn Gly Ser Thr Val Pro SerPro Val Lys Asp 195 200 205 Ile Gly Ile Ser Thr Lys Ala Asn Ser Leu LeuIle Ser Trp Ser His 210 215 220 Gly Ser Gly Asn Val Glu Arg Tyr Arg LeuMet Leu Met Asp Lys Gly 225 230 235 240 Ile Leu Val His Gly Gly Val ValAsp Lys His Ala Thr Ser Tyr Ala 245 250 255 Phe His Gly Leu Ser Pro GlyTyr Leu Tyr Asn Leu Thr Val Met Thr 260 265 270 Glu Ala Ala Gly Leu GlnAsn Tyr Arg Trp Lys Leu Val Arg Thr Ala 275 280 285 Pro Met Glu Val SerAsn Leu Lys Val Thr Asn Asp Gly Ser Leu Thr 290 295 300 Ser Leu Lys ValLys Trp Gln Arg Pro Pro Gly Asn Val Asp Ser Tyr 305 310 315 320 Asn IleThr Leu Ser His Lys Gly Thr Ile Lys Glu Ser Arg Val Leu 325 330 335 AlaPro Trp Ile Thr Glu Thr His Phe Lys Glu Leu Val Pro Gly Arg 340 345 350Leu Tyr Gln Val Thr Val Ser Cys Val Ser Gly Glu Leu Ser Ala Gln 355 360365 Lys Met Ala Val Gly Arg Thr Phe Pro Asp Lys Val Ala Asn Leu Glu 370375 380 Ala Asn Asn Asn Gly Arg Met Arg Ser Leu Val Val Ser Trp Ser Pro385 390 395 400 Pro Ala Gly Asp Trp Glu Gln Tyr Arg Ile Leu Leu Phe AsnAsp Ser 405 410 415 Val Val Leu Leu Asn Ile Thr Val Gly Lys Glu Glu ThrGln Tyr Val 420 425 430 Met Asp Asp Thr Gly Leu Val Pro Gly Arg Gln TyrGlu Val Glu Val 435 440 445 Ile Val Glu Ser Gly Asn Leu Lys Asn Ser GluArg Cys Gln Gly Arg 450 455 460 Thr Val Pro Leu Ala Val Leu Gln Leu ArgVal Lys His Ala Asn Glu 465 470 475 480 Thr Ser Leu Ser Ile Met Trp GlnThr Pro Val Ala Glu Trp Glu Lys 485 490 495 Tyr Ile Ile Ser Leu Ala AspArg Asp Leu Leu Leu Ile His Lys Ser 500 505 510 Leu Ser Lys Asp Ala LysGlu Phe Thr Phe Thr Asp Leu Val Pro Gly 515 520 525 Arg Lys Tyr Met AlaThr Val Thr Ser Ile Ser Gly Asp Leu Lys Asn 530 535 540 Ser Ser Ser ValLys Gly Arg Thr Val Pro Ala Gln Val Thr Asp Leu 545 550 555 560 His ValAla Asn Gln Gly Met Thr Ser Ser Leu Phe Thr Asn Trp Thr 565 570 575 GlnAla Gln Gly Asp Val Glu Phe Tyr Gln Val Leu Leu Ile His Glu 580 585 590Asn Val Val Ile Lys Asn Glu Ser Ile Ser Ser Glu Thr Ser Arg Tyr 595 600605 Ser Phe His Ser Leu Lys Ser Gly Ser Leu Tyr Ser Val Val Val Thr 610615 620 Thr Val Ser Gly Gly Ile Ser Ser Arg Gln Val Val Val Glu Gly Arg625 630 635 640 Thr Val Pro Ser Ser Val Ser Gly Val Thr Val Asn Asn SerGly Arg 645 650 655 Asn Asp Tyr Leu Ser Val Ser Trp Leu Val Ala Pro GlyAsp Val Asp 660 665 670 Asn Tyr Glu Val Thr Leu Ser His Asp Gly Lys ValVal Gln Ser Leu 675 680 685 Val Ile Ala Lys Ser Val Arg Glu Cys Ser PheSer Ser Leu Thr Pro 690 695 700 Gly Arg Leu Tyr Thr Val Thr Ile Thr ThrArg Ser Gly Lys Tyr Glu 705 710 715 720 Asn His Ser Phe Ser Gln Glu ArgThr Val Pro Asp Lys Val Gln Gly 725 730 735 Val Ser Val Ser Asn Ser AlaArg Ser Asp Tyr Leu Arg Val Ser Trp 740 745 750 Val Tyr Ala Thr Gly AspPhe Asp His Tyr Glu Val Thr Ile Lys Asn 755 760 765 Lys Asn Asn Phe IleGln Thr Lys Ser Ile Pro Lys Ser Glu Asn Glu 770 775 780 Cys Val Phe ValGln Leu Val Pro Gly Arg Leu Tyr Ser Val Thr Val 785 790 795 800 Thr ThrLys Ser Gly Gln Tyr Glu Ala Asn Glu Gln Gly Asn Gly Arg 805 810 815 ThrIle Pro Glu Pro Val Lys Asp Leu Thr Leu Arg Asn Arg Ser Thr 820 825 830Glu Asp Leu His Val Thr Trp Ser Gly Ala Asn Gly Asp Val Asp Gln 835 840845 Tyr Glu Ile Gln Leu Leu Phe Asn Asp Met Lys Val Phe Pro Pro Phe 850855 860 His Leu Val Asn Thr Ala Thr Glu Tyr Arg Phe Thr Ser Leu Thr Pro865 870 875 880 Gly Arg Gln Tyr Lys Ile Leu Val Leu Thr Ile Ser Gly AspVal Gln 885 890 895 Gln Ser Ala Phe Ile Glu Gly Phe Thr Val Pro Ser AlaVal Lys Asn 900 905 910 Ile His Ile Ser Pro Asn Gly Ala Thr Asp Ser LeuThr Val Asn Trp 915 920 925 Thr Pro Gly Gly Gly Asp Val Asp Ser Tyr ThrVal Ser Ala Phe Arg 930 935 940 His Ser Gln Lys Val Asp Ser Gln Thr IlePro Lys His Val Phe Glu 945 950 955 960 His Thr Phe His Arg Leu Glu AlaGly Glu Gln Tyr Gln Ile Met Ile 965 970 975 Ala Ser Val Ser Gly Ser LeuLys Asn Gln Ile Asn Val Val Gly Arg 980 985 990 Thr Val Pro Ala Ser ValGln Gly Val Ile Ala Asp Asn Ala Tyr Ser 995 1000 1005 Ser Tyr Ser LeuIle Val Ser Trp Gln Lys Ala Ala Gly Val Ala 1010 1015 1020 Glu Arg TyrAsp Ile Leu Leu Leu Thr Glu Asn Gly Ile Leu Leu 1025 1030 1035 Arg AsnThr Ser Glu Pro Ala Thr Thr Lys Gln His Lys Phe Glu 1040 1045 1050 AspLeu Thr Pro Gly Lys Lys Tyr Lys Ile Gln Ile Leu Thr Val 1055 1060 1065Ser Gly Gly Leu Phe Ser Lys Glu Ala Gln Thr Glu Gly Arg Thr 1070 10751080 Val Pro Ala Ala Val Thr Asp Leu Arg Ile Thr Glu Asn Ser Thr 10851090 1095 Arg His Leu Ser Phe Arg Trp Thr Ala Ser Glu Gly Glu Leu Ser1100 1105 1110 Trp Tyr Asn Ile Phe Leu Tyr Asn Pro Asp Gly Asn Leu GlnGlu 1115 1120 1125 Arg Ala Gln Val Asp Pro Leu Val Gln Ser Phe Ser PheGln Asn 1130 1135 1140 Leu Leu Gln Gly Arg Met Tyr Lys Met Val Ile ValThr His Ser 1145 1150 1155 Gly Glu Leu Ser Asn Glu Ser Phe Ile Phe GlyArg Thr Val Pro 1160 1165 1170 Ala Ser Val Ser His Leu Arg Gly Ser AsnArg Asn Thr Thr Asp 1175 1180 1185 Ser Leu Trp Phe Asn Trp Ser Pro AlaSer Gly Asp Phe Asp Phe 1190 1195 1200 Tyr Glu Leu Ile Leu Tyr Asn ProAsn Gly Thr Lys Lys Glu Asn 1205 1210 1215 Trp Lys Asp Lys Asp Leu ThrGlu Trp Arg Phe Gln Gly Leu Val 1220 1225 1230 Pro Gly Arg Lys Tyr ValLeu Trp Val Val Thr His Ser Gly Asp 1235 1240 1245 Leu Ser Asn Lys ValThr Ala Glu Ser Arg Thr Ala Pro Ser Pro 1250 1255 1260 Pro Ser Leu MetSer Phe Ala Asp Ile Ala Asn Thr Ser Leu Ala 1265 1270 1275 Ile Thr TrpLys Gly Pro Pro Asp Trp Thr Asp Tyr Asn Asp Phe 1280 1285 1290 Glu LeuGln Trp Leu Pro Arg Asp Ala Leu Thr Val Phe Asn Pro 1295 1300 1305 TyrAsn Asn Arg Lys Ser Glu Gly Arg Ile Val Tyr Gly Leu Arg 1310 1315 1320Pro Gly Arg Ser Tyr Gln Phe Asn Val Lys Thr Val Ser Gly Asp 1325 13301335 Ser Trp Lys Thr Tyr Ser Lys Pro Ile Phe Gly Ser Val Arg Thr 13401345 1350 Lys Pro Asp Lys Ile Gln Asn Leu His Cys Arg Pro Gln Asn Ser1355 1360 1365 Thr Ala Ile Ala Cys Ser Trp Ile Pro Pro Asp Ser Asp PheAsp 1370 1375 1380 Gly Tyr Ser Ile Glu Cys Arg Lys Met Asp Thr Gln GluVal Glu 1385 1390 1395 Phe Ser Arg Lys Leu Glu Lys Glu Lys Ser Leu LeuAsn Ile Met 1400 1405 1410 Met Leu Val Pro His Lys Arg Tyr Leu Val SerIle Lys Val Gln 1415 1420 1425 Ser Ala Gly Met Thr Ser Glu Val Val GluAsp Ser Thr Ile Thr 1430 1435 1440 Met Ile Asp Arg Pro Pro Pro Pro ProPro His Ile Arg Val Asn 1445 1450 1455 Glu Lys Asp Val Leu Ile Ser LysSer Ser Ile Asn Phe Thr Val 1460 1465 1470 Asn Cys Ser Trp Phe Ser AspThr Asn Gly Ala Val Lys Tyr Phe 1475 1480 1485 Thr Val Val Val Arg GluAla Asp Gly Ser Asp Glu Leu Lys Pro 1490 1495 1500 Glu Gln Gln His ProLeu Pro Ser Tyr Leu Glu Tyr Arg His Asn 1505 1510 1515 Ala Ser Ile ArgVal Tyr Gln Thr Asn Tyr Phe Ala Ser Lys Cys 1520 1525 1530 Ala Glu AsnPro Asn Ser Asn Ser Lys Ser Phe Asn Ile Lys Leu 1535 1540 1545 Gly AlaGlu Met Glu Ser Leu Gly Gly Lys Cys Asp Pro Thr Gln 1550 1555 1560 GlnLys Phe Cys Asp Gly Pro Leu Lys Pro His Thr Ala Tyr Arg 1565 1570 1575Ile Ser Ile Arg Ala Phe Thr Gln Leu Phe Asp Glu Asp Leu Lys 1580 15851590 Glu Phe Thr Lys Pro Leu Tyr Ser Asp Thr Phe Phe Ser Leu Pro 15951600 1605 Ile Thr Thr Glu Ser Glu Pro Leu Phe Gly Ala Ile Glu Gly Val1610 1615 1620 Ser Ala Gly Leu Phe Leu Ile Gly Met Leu Val Ala Val ValAla 1625 1630 1635 Leu Leu Ile Cys Arg Gln Lys Val Ser His Gly Arg GluArg Pro 1640 1645 1650 Ser Ala Arg Leu Ser Ile Arg Arg Asp Arg Pro LeuSer Val His 1655 1660 1665 Leu Asn Leu Gly Gln Lys Gly Asn Arg Lys ThrSer Cys Pro Ile 1670 1675 1680 Lys Ile Asn Gln Phe Glu Gly His Phe MetLys Leu Gln Ala Asp 1685 1690 1695 Ser Asn Tyr Leu Leu Ser Lys Glu TyrGlu Glu Leu Lys Asp Val 1700 1705 1710 Gly Arg Asn Gln Ser Cys Asp IleAla Leu Leu Pro Glu Asn Arg 1715 1720 1725 Gly Lys Asn Arg Tyr Asn AsnIle Leu Pro Tyr Asp Ala Thr Arg 1730 1735 1740 Val Lys Leu Ser Asn ValAsp Asp Asp Pro Cys Ser Asp Tyr Ile 1745 1750 1755 Asn Ala Ser Tyr IlePro Gly Asn Asn Phe Arg Arg Glu Tyr Ile 1760 1765 1770 Val Thr Gln GlyPro Leu Pro Gly Thr Lys Asp Asp Phe Trp Lys 1775 1780 1785 Met Val TrpGlu Gln Asn Val His Asn Ile Val Met Val Thr Gln 1790 1795 1800 Cys ValGlu Lys Gly Arg Val Lys Cys Asp His Tyr Trp Pro Ala 1805 1810 1815 AspGln Asp Ser Leu Tyr Tyr Gly Asp Leu Ile Leu Gln Met Leu 1820 1825 1830Ser Glu Ser Val Leu Pro Glu Trp Thr Ile Arg Glu Phe Lys Ile 1835 18401845 Cys Gly Glu Glu Gln Leu Asp Ala His Arg Leu Ile Arg His Phe 18501855 1860 His Tyr Thr Val Trp Pro Asp His Gly Val Pro Glu Thr Thr Gln1865 1870 1875 Ser Leu Ile Gln Phe Val Arg Thr Val Arg Asp Tyr Ile AsnArg 1880 1885 1890 Ser Pro Gly Ala Gly Pro Thr Val Val His Cys Ser AlaGly Val 1895 1900 1905 Gly Arg Thr Gly Thr Phe Ile Ala Leu Asp Arg IleLeu Gln Gln 1910 1915 1920 Leu Asp Ser Lys Asp Ser Val Asp Ile Tyr GlyAla Val His Asp 1925 1930 1935 Leu Arg Leu His Arg Val His Met Val GlnThr Glu Cys Gln Tyr 1940 1945 1950 Val Tyr Leu His Gln Cys Val Arg AspVal Leu Arg Ala Arg Lys 1955 1960 1965 Leu Arg Ser Glu Gln Glu Asn ProLeu Phe Pro Ile Tyr Glu Asn 1970 1975 1980 Val Asn Pro Glu Tyr His ArgAsp Pro Val Tyr Ser Arg His 1985 1990 1995 55 453 PRT Homo sapien 55 MetLys Leu Leu Val Ile Leu Leu Phe Ser Gly Leu Ile Thr Gly Phe 1 5 10 15Arg Ser Asp Ser Ser Ser Ser Leu Pro Pro Lys Leu Leu Leu Val Ser 20 25 30Phe Asp Gly Phe Arg Ala Asp Tyr Leu Lys Asn Tyr Glu Phe Pro His 35 40 45Leu Gln Asn Phe Ile Lys Glu Gly Val Leu Val Glu His Val Lys Asn 50 55 60Val Phe Ile Thr Lys Thr Phe Pro Asn His Tyr Ser Ile Val Thr Gly 65 70 7580 Leu Tyr Glu Glu Ser His Gly Ile Val Ala Asn Ser Met Tyr Asp Ala 85 9095 Val Thr Lys Lys His Phe Ser Asp Ser Asn Asp Lys Asp Pro Phe Trp 100105 110 Trp Asn Glu Ala Val Pro Ile Trp Val Thr Asn Gln Leu Gln Glu Asn115 120 125 Arg Ser Ser Ala Ala Ala Met Trp Pro Gly Thr Asp Val Pro IleHis 130 135 140 Asp Thr Ile Ser Ser Tyr Phe Met Asn Tyr Asn Ser Ser ValSer Phe 145 150 155 160 Glu Glu Arg Leu Asn Asn Ile Thr Met Trp Leu AsnAsn Ser Asn Pro 165 170 175 Pro Val Thr Phe Ala Thr Leu Tyr Trp Glu GluPro Asp Ala Ser Gly 180 185 190 His Lys Tyr Gly Pro Glu Asp Lys Glu AsnMet Ser Arg Val Leu Lys 195 200 205 Lys Ile Asp Asp Leu Ile Gly Asp LeuVal Gln Arg Leu Lys Met Leu 210 215 220 Gly Leu Trp Glu Asn Leu Asn ValIle Ile Thr Ser Asp His Gly Met 225 230 235 240 Thr Gln Cys Ser Gln AspArg Leu Ile Asn Leu Asp Ser Cys Ile Asp 245 250 255 His Ser Tyr Tyr ThrLeu Ile Asp Leu Ser Pro Val Ala Ala Ile Leu 260 265 270 Pro Lys Ile AsnArg Thr Glu Val Tyr Asn Lys Leu Lys Asn Cys Ser 275 280 285 Pro His MetAsn Val Tyr Leu Lys Glu Asp Ile Pro Asn Arg Phe Tyr 290 295 300 Tyr GlnHis Asn Asp Arg Ile Gln Pro Ile Ile Leu Val Ala Asp Glu 305 310 315 320Gly Trp Thr Ile Val Leu Asn Glu Ser Ser Gln Lys Leu Gly Asp His 325 330335 Gly Tyr Asp Asn Ser Leu Pro Ser Met His Pro Phe Leu Ala Ala His 340345 350 Gly Pro Ala Phe His Lys Gly Tyr Lys His Ser Thr Ile Asn Ile Val355 360 365 Asp Ile Tyr Pro Met Met Cys His Ile Leu Gly Leu Lys Pro HisPro 370 375 380 Asn Asn Gly Thr Phe Gly His Thr Lys Cys Leu Leu Val AspGln Trp 385 390 395 400 Cys Ile Asn Leu Pro Glu Ala Ile Ala Ile Val IleGly Ser Leu Leu 405 410 415 Val Leu Thr Met Leu Thr Cys Leu Ile Ile IleMet Gln Asn Arg Leu 420 425 430 Ser Val Pro Arg Pro Phe Ser Arg Leu GlnLeu Gln Glu Asp Asp Asp 435 440 445 Asp Pro Leu Ile Gly 450 56 537 PRTHomo sapien 56 Met Ser Lys Pro His Ser Glu Ala Gly Thr Ala Phe Ile GlnThr Gln 1 5 10 15 Gln Leu His Ala Ala Met Ala Asp Thr Phe Leu Glu HisMet Cys Arg 20 25 30 Leu Asp Ile Asp Ser Pro Pro Ile Thr Ala Arg Asn ThrGly Ile Ile 35 40 45 Cys Thr Ile Gly Pro Ala Ser Arg Ser Val Glu Thr LeuLys Glu Met 50 55 60 Ile Lys Ser Gly Met Asn Val Ala Arg Leu Asn Phe SerHis Gly Thr 65 70 75 80 His Glu Tyr His Ala Glu Thr Ile Lys Asn Val ArgThr Ala Thr Glu 85 90 95 Ser Phe Ala Ser Asp Pro Ile Leu Tyr Arg Pro ValAla Val Ala Leu 100 105 110 Asp Thr Lys Gly Pro Glu Ile Arg Thr Gly LeuIle Lys Gly Ser Gly 115 120 125 Thr Ala Glu Val Glu Leu Lys Lys Gly AlaThr Leu Lys Ile Thr Leu 130 135 140 Asp Asn Ala Tyr Met Glu Lys Cys AspGlu Asn Ile Leu Trp Leu Asp 145 150 155 160 Tyr Lys Asn Ile Cys Lys ValVal Glu Val Gly Ser Lys Ile Tyr Val 165 170 175 Asp Asp Gly Leu Ile SerLeu Gln Val Lys Gln Lys Gly Ala Asp Phe 180 185 190 Leu Val Thr Glu ValGlu Asn Gly Gly Ser Leu Gly Ser Lys Lys Gly 195 200 205 Val Asn Leu ProGly Ala Ala Val Asp Leu Pro Ala Val Ser Glu Lys 210 215 220 Asp Ile ProGly Ser Glu Ser Leu Gly Val Glu Gln Asp Val Asp Met 225 230 235 240 ValPhe Ala Ser Phe His Pro Ala Lys Ala Ser Gly Cys Pro Met Glu 245 250 255Ala Leu Gly Ala Val Leu Gly Arg Glu Gly Lys Arg Asn Ile Lys Ile 260 265270 Ile Ser Lys Ile Glu Asn His Glu Gly Val Arg Arg Phe Asp Glu Ile 275280 285 Leu Glu Ala Ser Asp Gly Ile Met Val Ala Arg Gly Asp Leu Gly Ile290 295 300 Glu Ile Pro Ala Glu Lys Val Phe Leu Ala Gln Lys Met Met IleGly 305 310 315 320 Arg Cys Asn Pro Arg Thr Gly Lys Pro Val Ile Cys AlaThr Gln Met 325 330 335 Leu Glu Ser Ile Ile Lys Lys Pro Arg Pro Thr ArgAla Glu Gly Ser 340 345 350 Asp Val Ala Asn Ala Val Leu Asp Gly Ala AspCys Ile Met Leu Ser 355 360 365 Gly Glu Thr Ala Lys Gly Asp Tyr Pro LeuGlu Ala Val Arg Met Gln 370 375 380 His Leu Ile Ala Arg Glu Ala Glu AlaAla Ile Tyr His Leu Gln Leu 385 390 395 400 Phe Glu Glu Leu Arg Arg LeuAla Pro Ile Thr Ser Asp Pro Thr Glu 405 410 415 Ala Thr Ala Val Gly AlaVal Glu Ala Ser Phe Lys Cys Cys Ser Gly 420 425 430 Ala Ile Ile Val LeuThr Lys Ser Gly Arg Ser Ala His Gln Val Ala 435 440 445 Arg Tyr Arg ProArg Ala Pro Ile Ile Ala Val Thr Arg Asn Pro Gln 450 455 460 Thr Ala ArgGln Ala His Leu Tyr Arg Gly Ile Phe Pro Val Leu Cys 465 470 475 480 LysAsp Pro Val Gln Glu Ala Trp Ala Glu Asp Val Asp Leu Arg Val 485 490 495Asn Phe Ala Met Asn Val Gly Lys Ala Arg Gly Phe Phe Lys Lys Gly 500 505510 Asp Val Val Ile Val Leu Thr Gly Trp Arg Pro Gly Ser Gly Phe Thr 515520 525 Asn Thr Met Arg Val Val Pro Val Pro 530 535

What is claimed is:
 1. A LSG comprising: (a) a polynucleotide of SEQ IDNO:1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19,20, 21, 22, 37 or 38 or a variant thereof; (b) a polypeptide expressedby a polynucleotide of SEQ ID NO:1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 37 or 38 or a variant thereof;or (c) a polynucleotide which is capable of hybridizing under stringentconditions to the antisense sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7,8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 37 or
 38. 2.The LSG of claim 1 wherein the polypeptide comprises SEQ ID NO: 39, 40,41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, or
 56. 3. Amethod for diagnosing the presence of lung cancer in a patientcomprising: (a) determining levels of a LSG of claim 1 in cells, tissuesor bodily fluids in a patient; and (b) comparing the determined levelsof LSG with levels of LSG in cells, tissues or bodily fluids from anormal human control, wherein a change in determined levels of LSG insaid patient versus normal human control is associated with the presenceof lung cancer.
 4. A method of diagnosing metastases of lung cancer in apatient comprising: (a) identifying a patient having lung cancer that isnot known to have metastasized; (b) determining levels of a LSG of claim1 in a sample of cells, tissues, or bodily fluid from said patient; and(c) comparing the determined LSG levels with levels of LSG in cells,tissue, or bodily fluid of a normal human control, wherein an increasein determined LSG levels in the patient versus the normal human controlis associated with a cancer which has metastasized.
 5. A method ofstaging lung cancer in a patient having lung cancer comprising: (a)identifying a patient having lung cancer; (b) determining levels of aLSG of claim 1 in a sample of cells, tissue, or bodily fluid from saidpatient; and (c) comparing determined LSG levels with levels of LSG incells, tissues, or bodily fluid of a normal human control, wherein anincrease in determined LSG levels in said patient versus the normalhuman control is associated with a cancer which is progressing and adecrease in the determined LSG levels is associated with a cancer whichis regressing or in remission.
 6. A method of monitoring lung cancer ina patient for the onset of metastasis comprising: (a) identifying apatient having lung cancer that is not known to have metastasized; (b)periodically determining levels of a LSG of claim 1 in samples of cells,tissues, or bodily fluid from said patient; and (c) comparing theperiodically determined LSG levels with levels of LSG in cells, tissues,or bodily fluid of a normal human control, wherein an increase in anyone of the periodically determined LSG levels in the patient versus thenormal human control is associated with a cancer which has metastasized.7. A method of monitoring a change in stage of lung cancer in a patientcomprising: (a) identifying a patient having lung cancer; (b)periodically determining levels of a LSG of claim 1 in cells, tissues,or bodily fluid from said patient; and (c) comparing the periodicallydetermined LSG levels with levels of LSG in cells, tissues, or bodilyfluid of a normal human control, wherein an increase in any one of theperiodically determined LSG levels in the patient versus the normalhuman control is associated with a cancer which is progressing in stageand a decrease is associated with a cancer which is regressing in stageor in remission.
 8. A method of identifying potential therapeutic agentsfor use in imaging and treating lung cancer comprising screeningcompounds for an ability to bind to or decrease expression of a LSG ofclaim 1 relative to the LSG in the absence of the compound wherein theability of the compound to bind to the LSG or decrease expression of theLSG is indicative of the compound being useful in imaging and treatinglung cancer.
 9. An antibody which specifically binds a polypeptideencoded by a LSG of claim
 1. 10. The antibody of claim 9 wherein thepolypeptide comprises SEQ ID NO:39, 40, 41, 42, 43, 44, 45, 46, 47, 48,49, 50, 51, 52, 53, 54, 55 or
 56. 11. A method of imaging lung cancer ina patient comprising administering to the patient an antibody of claim9.
 12. The method of claim 11 wherein said antibody is labeled withparamagnetic ions or a radioisotope.
 13. A method of treating lungcancer in a patient comprising administering to the patient a compoundwhich downregulates expression or activity of a LSG of claim
 1. 14. Amethod of inducing an immune response against a target cell expressing aLSG of claim 1 comprising delivering to a human patient animmunogenically stimulatory amount of a LSG polypeptide so that animmune response is mounted against the target cell.
 15. The method ofclaim 14 wherein the LSG polypeptide comprises SEQ ID NO:39, 40, 41, 42,43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55 or
 56. 16. A vaccinefor treating lung cancer comprising a LSG of claim 1.